                         SEQUENCE LISTING

<110>  Codexis, Inc.
 
<120>  ENGINEERED LIGASE VARIANTS

<130>  CX9-160WO1

<150>  US 62/503075
<151>  2017-05-08

<150>  US 62/540734
<151>  2017-08-03

<150>  US 62/587030
<151>  2017-11-16

<160>  54    

<170>  PatentIn version 3.5

<210>  1
<211>  1464
<212>  DNA
<213>  Bacteriophage T4

<400>  1
atgattctta aaattctgaa cgaaatagca tctattggtt caactaaaca gaagcaagca       60

attcttgaaa agaataaaga taatgaattg cttaaacgag tatatcgtct gacttattct      120

cgtgggttac agtattatat caagaaatgg cctaaacctg gtattgctac ccagagtttt      180

ggaatgttga ctcttaccga tatgcttgac ttcattgaat tcacattagc tactcggaaa      240

ttgactggaa atgcagcaat tgaggaatta actggatata tcaccgatgg taaaaaagat      300

gatgttgaag ttttgcgtcg agtgatgatg cgagaccttg aatgtggtgc ttcagtatct      360

attgcaaaca aagtttggcc aggtttaatt cctgaacaac ctcaaatgct cgcaagttct      420

tatgatgaaa aaggcattaa taagaatatc aaatttccag cctttgctca gttaaaagct      480

gatggagctc ggtgttttgc tgaagttaga ggtgatgaat tagatgatgt tcgtctttta      540

tcacgagctg gtaatgaata tctaggatta gatcttctta aggaagagtt aattaaaatg      600

accgctgaag cccgccagat tcatccagaa ggtgtgttga ttgatggcga attggtatac      660

catgagcaag ttaaaaagga gccagaaggc ctagattttc tttttgatgc ttatcctgaa      720

aacagtaaag ctaaagaatt cgccgaagta gctgaatcac gtactgcttc taatggaatc      780

gccaataaat ctttaaaggg aaccatttct gaaaaagaag cacaatgcat gaagtttcag      840

gtctgggatt atgtcccgtt ggtagaaata tacagtcttc ctgcatttcg tttgaaatat      900

gatgtacgtt tttctaaact agaacaaatg acatctggat atgataaagt aattttaatt      960

gaaaaccagg tagtaaataa cctagatgaa gctaaggtaa tttataaaaa gtatattgac     1020

caaggtcttg aaggtattat tctcaaaaat atcgatggat tatgggaaaa tgctcgttca     1080

aaaaatcttt ataaatttaa agaagtaatt gatgttgatt taaaaattgt aggaatttat     1140

cctcaccgta aagaccctac taaagcgggt ggatttattc ttgagtcaga gtgtggaaaa     1200

attaaggtaa atgctggttc aggcttaaaa gataaagccg gtgtaaaatc gcatgaactt     1260

gaccgtactc gcattatgga aaaccaaaat tattatattg gaaaaattct agagtgcgaa     1320

tgcaacggtt ggttaaaatc tgatggccgc actgattacg ttaaattatt tcttccgatt     1380

gcgattcgtt tacgtgaaga taaaactaaa gctaatacat tcgaagatgt atttggtgat     1440

tttcatgagg taactggtct ataa                                            1464


<210>  2
<211>  487
<212>  PRT
<213>  Bacteriophage T4

<400>  2

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Glu Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Pro Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Lys Lys Glu Pro Glu Gly Leu Asp Phe Leu Phe Asp Ala Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Glu Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Thr Asp Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  3
<211>  1497
<212>  DNA
<213>  Bacteriophage T4

<400>  3
atgcatcacc atcaccatca cggtggcagc ggtatgattc ttaaaattct gaacgaaata       60

gcatctattg gttcaactaa acagaagcaa gcaattcttg aaaagaataa agataatgaa      120

ttgcttaaac gagtatatcg tctgacttat tctcgtgggt tacagtatta tatcaagaaa      180

tggcctaaac ctggtattgc tacccagagt tttggaatgt tgactcttac cgatatgctt      240

gacttcattg aattcacatt agctactcgg aaattgactg gaaatgcagc aattgaggaa      300

ttaactggat atatcaccga tggtaaaaaa gatgatgttg aagttttgcg tcgagtgatg      360

atgcgagacc ttgaatgtgg tgcttcagta tctattgcaa acaaagtttg gccaggttta      420

attcctgaac aacctcaaat gctcgcaagt tcttatgatg aaaaaggcat taataagaat      480

atcaaatttc cagcctttgc tcagttaaaa gctgatggag ctcggtgttt tgctgaagtt      540

agaggtgatg aattagatga tgttcgtctt ttatcacgag ctggtaatga atatctagga      600

ttagatcttc ttaaggaaga gttaattaaa atgaccgctg aagcccgcca gattcatcca      660

gaaggtgtgt tgattgatgg cgaattggta taccatgagc aagttaaaaa ggagccagaa      720

ggcctagatt ttctttttga tgcttatcct gaaaacagta aagctaaaga attcgccgaa      780

gtagctgaat cacgtactgc ttctaatgga atcgccaata aatctttaaa gggaaccatt      840

tctgaaaaag aagcacaatg catgaagttt caggtctggg attatgtccc gttggtagaa      900

atatacagtc ttcctgcatt tcgtttgaaa tatgatgtac gtttttctaa actagaacaa      960

atgacatctg gatatgataa agtaatttta attgaaaacc aggtagtaaa taacctagat     1020

gaagctaagg taatttataa aaagtatatt gaccaaggtc ttgaaggtat tattctcaaa     1080

aatatcgatg gattatggga aaatgctcgt tcaaaaaatc tttataaatt taaagaagta     1140

attgatgttg atttaaaaat tgtaggaatt tatcctcacc gtaaagaccc tactaaagcg     1200

ggtggattta ttcttgagtc agagtgtgga aaaattaagg taaatgctgg ttcaggctta     1260

aaagataaag ccggtgtaaa atcgcatgaa cttgaccgta ctcgcattat ggaaaaccaa     1320

aattattata ttggaaaaat tctagagtgc gaatgcaacg gttggttaaa atctgatggc     1380

cgcactgatt acgttaaatt atttcttccg attgcgattc gtttacgtga agataaaact     1440

aaagctaata cattcgaaga tgtatttggt gattttcatg aggtaactgg tctataa        1497


<210>  4
<211>  498
<212>  PRT
<213>  Bacteriophage T4

<400>  4

Met His His His His His His Gly Gly Ser Gly Met Ile Leu Lys Ile 
1               5                   10                  15      


Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys Gln Lys Gln Ala Ile 
            20                  25                  30          


Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys Arg Val Tyr Arg Leu 
        35                  40                  45              


Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys Lys Trp Pro Lys Pro 
    50                  55                  60                  


Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr Leu Thr Asp Met Leu 
65                  70                  75                  80  


Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys Leu Thr Gly Asn Ala 
                85                  90                  95      


Ala Ile Glu Glu Leu Thr Gly Tyr Ile Thr Asp Gly Lys Lys Asp Asp 
            100                 105                 110         


Val Glu Val Leu Arg Arg Val Met Met Arg Asp Leu Glu Cys Gly Ala 
        115                 120                 125             


Ser Val Ser Ile Ala Asn Lys Val Trp Pro Gly Leu Ile Pro Glu Gln 
    130                 135                 140                 


Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys Gly Ile Asn Lys Asn 
145                 150                 155                 160 


Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala Asp Gly Ala Arg Cys 
                165                 170                 175     


Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp Val Arg Leu Leu Ser 
            180                 185                 190         


Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu Leu Lys Glu Glu Leu 
        195                 200                 205             


Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His Pro Glu Gly Val Leu 
    210                 215                 220                 


Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val Lys Lys Glu Pro Glu 
225                 230                 235                 240 


Gly Leu Asp Phe Leu Phe Asp Ala Tyr Pro Glu Asn Ser Lys Ala Lys 
                245                 250                 255     


Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala Ser Asn Gly Ile Ala 
            260                 265                 270         


Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys Glu Ala Gln Cys Met 
        275                 280                 285             


Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val Glu Ile Tyr Ser Leu 
    290                 295                 300                 


Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe Ser Lys Leu Glu Gln 
305                 310                 315                 320 


Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile Glu Asn Gln Val Val 
                325                 330                 335     


Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys Lys Tyr Ile Asp Gln 
            340                 345                 350         


Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp Gly Leu Trp Glu Asn 
        355                 360                 365             


Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu Val Ile Asp Val Asp 
    370                 375                 380                 


Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys Asp Pro Thr Lys Ala 
385                 390                 395                 400 


Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys Ile Lys Val Asn Ala 
                405                 410                 415     


Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys Ser His Glu Leu Asp 
            420                 425                 430         


Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr Ile Gly Lys Ile Leu 
        435                 440                 445             


Glu Cys Glu Cys Asn Gly Trp Leu Lys Ser Asp Gly Arg Thr Asp Tyr 
    450                 455                 460                 


Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu Arg Glu Asp Lys Thr 
465                 470                 475                 480 


Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp Phe His Glu Val Thr 
                485                 490                 495     


Gly Leu 
        


<210>  5
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic T4 DNA ligase

<400>  5
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgctga ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tgaagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggcc gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttaaaaagga gccggagggg ttggatttcc tgtttgatgc ctacccggag      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt gatgtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggaatgcgag     1320

tgcaacgggt ggcttaagag cgacgggcgc acggactatg ttaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  6
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic T4 DNA ligase

<400>  6

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Glu Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Pro Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Lys Lys Glu Pro Glu Gly Leu Asp Phe Leu Phe Asp Ala Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Glu Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Thr Asp Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  7
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Artificial Oligonucleotides

<400>  7
tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcaca                  50


<210>  8
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  8
tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcaa                   49


<210>  9
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides


<220>
<221>  misc_feature
<222>  (20)..(20)
<223>  n is a, c, g, or t

<400>  9
acacgacgct cttccgatcn t                                                 21


<210>  10
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  10
gatcggaaga gcgtcgtgt                                                    19


<210>  11
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  11
tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcaca                  50


<210>  12
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  12
gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcaa                  50


<210>  13
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides


<220>
<221>  misc_feature
<222>  (20)..(20)
<223>  n is a, c, g, or t

<400>  13
acacgacgct cttccgatcn t                                                 21


<210>  14
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  14
gatcggaaga gcgtcgtgt                                                    19


<210>  15
<211>  65
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides


<220>
<221>  misc_feature
<222>  (64)..(64)
<223>  n is a, c, g, or t

<400>  15
gatcggaaga gcacacgtct gaactccagt cacactcttt ccctacacga cgctcttccg       60

atcnt                                                                   65


<210>  16
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  16
gatcggaaga gcgtcgtgt                                                    19


<210>  17
<211>  59
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides


<220>
<221>  misc_feature
<222>  (58)..(58)
<223>  n is a, c, g, or t

<400>  17
aatgatacgg cgaccaccga gatctacact ctttccctac acgacgctct tccgatcnt        59


<210>  18
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  18
gatcggaaga gcacacgtct gaactccagt cacatcacga tctcgtatgc cgtcttctgc       60

ttg                                                                     63


<210>  19
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  19
aagccggtgt aaaatcgcat ga                                                22


<210>  20
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  20
agtaaacgaa tcgcaatcgg aaga                                              24


<210>  21
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  21
cagccggtgt aaaatcgcat ga                                                22


<210>  22
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  22
cgtaaacgaa tcgcaatcgg aaga                                              24


<210>  23
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  23
gagccggtgt aaaatcgcat ga                                                22


<210>  24
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  24
ggtaaacgaa tcgcaatcgg aaga                                              24


<210>  25
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  25
tagccggtgt aaaatcgcat ga                                                22


<210>  26
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  26
tgtaaacgaa tcgcaatcgg aaga                                              24


<210>  27
<211>  160
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  27
aagccggtgt aaaatcgcat gaacttgacc gtactcgcat tatggaaaac caaaattatt       60

atattggaaa aattctagag tgcgaatgca acggttggtt aaaatctgat ggccgcactg      120

attacgttaa attatttctt ccgattgcga ttcgtttaca                            160


<210>  28
<211>  160
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  28
cagccggtgt aaaatcgcat gaacttgacc gtactcgcat tatggaaaac caaaattatt       60

atattggaaa aattctagag tgcgaatgca acggttggtt aaaatctgat ggccgcactg      120

attacgttaa attatttctt ccgattgcga ttcgtttacc                            160


<210>  29
<211>  160
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  29
gagccggtgt aaaatcgcat gaacttgacc gtactcgcat tatggaaaac caaaattatt       60

atattggaaa aattctagag tgcgaatgca acggttggtt aaaatctgat ggccgcactg      120

attacgttaa attatttctt ccgattgcga ttcgtttacg                            160


<210>  30
<211>  160
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic Oligonucleotides

<400>  30
tagccggtgt aaaatcgcat gaacttgacc gtactcgcat tatggaaaac caaaattatt       60

atattggaaa aattctagag tgcgaatgca acggttggtt aaaatctgat ggccgcactg      120

attacgttaa attatttctt ccgattgcga ttcgtttact                            160


<210>  31
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  31
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgctga ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggcc gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatttcc tgtttgatgc ctacccggag      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt gatgtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaagactatg ttaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  32
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  32

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Pro Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Phe Leu Phe Asp Ala Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Asp Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  33
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  33
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgctga ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatttcc tgtttgatgc ctacccggag      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt gatgtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaagactatg ttaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  34
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  34

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Phe Leu Phe Asp Ala Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Asp Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  35
<211>  1467
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  35
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaagaatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatttcc tgtttgatgc ttacccgcct      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaaccctatg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcactggtct gtaataa                                         1467


<210>  36
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  36

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Arg Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Phe Leu Phe Asp Ala Tyr Pro Pro 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Trp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Pro Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  37
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  37
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaaaaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatgctc tgtttgatcg atacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaaccctatg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  38
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  38

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Lys Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Ala Leu Phe Asp Arg Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Trp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Pro Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  39
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  39
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgctta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagaaaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatgctc tgtttgatcg atacccgcct      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt gatgtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggagtgcaaa     1320

tgcaacgggt ggcttaagag cgctgggcgc aaagacggtc tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  40
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  40

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Lys Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Ala Leu Phe Asp Arg Tyr Pro Pro 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Ala 
        435                 440                 445             


Gly Arg Lys Asp Gly Leu Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  41
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  41
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgctta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaagaatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatgctc tgtttgataa ttacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt gatgtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggagtgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaagactatc tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  42
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  42

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Arg Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Ala Leu Phe Asp Asn Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Asp Tyr Leu Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  43
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  43
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaacaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgctga ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctcggat tagaaaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaagaatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatgctc tgtttgataa ttacccgcct      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt gatgtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacccgac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga aaaccagaac tattatatcg gcaaaattct ggagtgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaagactatg ttaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  44
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  44

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Gln Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Leu Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Arg Ile Arg Lys Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Arg Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Ala Leu Phe Asp Asn Tyr Pro Pro 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Asp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Asp Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  45
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  45
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaaaaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaacatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcaaag gtaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg aaagacgctc tgtttgatcg atacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagaccctac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgcg gaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aagccctatg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcggaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  46
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  46

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Lys Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Lys Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Lys Asp Ala Leu Phe Asp Arg Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Trp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Arg Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Pro Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Gly Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  47
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  47
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaaaaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcgctg gcaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatgctc tgtttgatcg atacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aaaaacctac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagcag gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga gaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aagccccgtg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcggaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  48
<211>  486
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  48

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Lys Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Leu 
    50                  55                  60                  


Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys Leu 
65                  70                  75                  80  


Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp Gly 
                85                  90                  95      


Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp Leu 
            100                 105                 110         


Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly Leu 
        115                 120                 125             


Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys Gly 
    130                 135                 140                 


Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala Asp 
145                 150                 155                 160 


Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp Val 
                165                 170                 175     


Arg Leu Leu Ser Arg Ala Gly Asn Glu Tyr Leu Gly Leu Asp Leu Leu 
            180                 185                 190         


Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His Pro 
        195                 200                 205             


Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val Ala 
    210                 215                 220                 


Lys Glu Pro Glu Gly Leu Asp Ala Leu Phe Asp Arg Tyr Pro Glu Asn 
225                 230                 235                 240 


Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala Ser 
                245                 250                 255     


Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys Glu 
            260                 265                 270         


Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val Glu 
        275                 280                 285             


Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe Ser 
    290                 295                 300                 


Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile Glu 
305                 310                 315                 320 


Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys Lys 
                325                 330                 335     


Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp Gly 
            340                 345                 350         


Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu Val 
        355                 360                 365             


Ile Trp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys Lys 
    370                 375                 380                 


Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys Ile 
385                 390                 395                 400 


Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys Ser 
                405                 410                 415     


His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr Ile 
            420                 425                 430         


Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp Gly 
        435                 440                 445             


Arg Lys Pro Arg Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu Arg 
    450                 455                 460                 


Gly Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp Phe 
465                 470                 475                 480 


His Glu Val Thr Gly Leu 
                485     


<210>  49
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  49
atgattctta aaattctgaa cgaaattgca agcattaagt ccactaaaca aaaaaaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaacatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcaaag gtaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttaaaagctc tgtttgatcg atacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagacaaaac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagctg gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga gaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc ggtccctatg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  50
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  50

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Lys Ser Thr Lys 
1               5                   10                  15      


Gln Lys Lys Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Lys Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Lys Ala Leu Phe Asp Arg Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Trp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Lys Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Gly Pro Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  51
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  51
atgattctta aaattctgaa cgaaattgca agcattggtt ccactaaaca aaaaaaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaatatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcaaag gtaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttggatgctc tgtttgatcg atacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctggatgaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagctc tgaagattgt tggtatttac     1140

ccgcatcgca aagacaaaac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagcag gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga gaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aaaccctatg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcgaaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  52
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  52

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Gly Ser Thr Lys 
1               5                   10                  15      


Gln Lys Lys Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Lys Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Asp Ala Leu Phe Asp Arg Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Asp Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Trp Val Ala Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Lys Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Pro Tyr Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Glu Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


<210>  53
<211>  1464
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  53
atgattctta aaattctgaa cgaaattgca agcattaagt ccactaaaca aaaaaaagcc       60

attctggaaa aaaataaaga caatgaactg ttaaagcgcg tgtatcgcct gacctattcg      120

cgtggcctgc aatactatat taaaaaatgg cccaaaccgg gcattgcgac gcagagcttc      180

ggcatgacta ccctgaccga tatgctggat tttatcgagt ttactcttgc gacgcgcaaa      240

cttaccggca acgctgctat tagagaattg acgggctata ttaccgatgg caaaaaagat      300

gatgttgagg tgctgcgtcg cgtcatgatg cgtgatctgg agtgcggtgc gtcagtgagc      360

atcgctaaca aagtgtggaa gggtttgatc ccggaacagc cacagatgct tgcaagcagc      420

tacgatgaaa agggcattaa caaaaacatc aaatttccgg ctttcgccca gctgaaagcg      480

gatggcgcgc gctgcttcgc cgaggtacgc ggtgacgaac tggacgacgt tcgtctgctg      540

tctcgcaaag gtaatgaata tctgggtctg gatctgctga aagaagaact gattaagatg      600

accgccgagg cgcgccaaat ccaccccgaa ggggtgctga ttgatggcga actggtgtat      660

cacgaacagg ttgcaaaaga gccggagggg ttaaaagctc tgtttgatcg atacccggaa      720

aatagcaaag cgaaagaatt tgcggaagtg gcggaatccc gcaccgcaag caatggtatc      780

gcaaataaat cgctgaaggg caccatttct gagaaagaag cacagtgtat gaagttccag      840

gtgtgggact acgtgccact ggttgagatc tactccctgc cagcgtttcg cctgaaatac      900

gatgtgcgtt tttcaaaact ggaacagatg acgagcggat atgataaagt aattttaatc      960

gagaaccagg tcgtgaacaa cctgctggaa gcaaaagtta tctataaaaa atatattgat     1020

cagggcttag aaggcattat cctgaagaac attgatggcc tttgggaaaa tgcacgcagc     1080

aaaaacctgt ataaatttaa agaagtgatt tgggtagatc tgaagattgt tggtatttac     1140

ccgcatcgca aagaccctac caaagcaggt ggtttcatcc tggaatctga atgcggtaaa     1200

attaaagtga acgcaggcag tggtttgaaa gataaagcag gtgttaaaag tcacgaactg     1260

gatcgcacgc gcatcatgga gaaccagaac tattatatcg gcaaaattct ggaatgcaaa     1320

tgcaacgggt ggcttaagag cgatgggcgc aagcccggtg tgaaattgtt cctgccgatt     1380

gcgattcgcc ttcgcggaga caaaactaag gcgaatactt tcgaagatgt gttcggtgat     1440

tttcatgaag tcacgggtct gtaa                                            1464


<210>  54
<211>  487
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  engineered ligase variant

<400>  54

Met Ile Leu Lys Ile Leu Asn Glu Ile Ala Ser Ile Lys Ser Thr Lys 
1               5                   10                  15      


Gln Lys Lys Ala Ile Leu Glu Lys Asn Lys Asp Asn Glu Leu Leu Lys 
            20                  25                  30          


Arg Val Tyr Arg Leu Thr Tyr Ser Arg Gly Leu Gln Tyr Tyr Ile Lys 
        35                  40                  45              


Lys Trp Pro Lys Pro Gly Ile Ala Thr Gln Ser Phe Gly Met Thr Thr 
    50                  55                  60                  


Leu Thr Asp Met Leu Asp Phe Ile Glu Phe Thr Leu Ala Thr Arg Lys 
65                  70                  75                  80  


Leu Thr Gly Asn Ala Ala Ile Arg Glu Leu Thr Gly Tyr Ile Thr Asp 
                85                  90                  95      


Gly Lys Lys Asp Asp Val Glu Val Leu Arg Arg Val Met Met Arg Asp 
            100                 105                 110         


Leu Glu Cys Gly Ala Ser Val Ser Ile Ala Asn Lys Val Trp Lys Gly 
        115                 120                 125             


Leu Ile Pro Glu Gln Pro Gln Met Leu Ala Ser Ser Tyr Asp Glu Lys 
    130                 135                 140                 


Gly Ile Asn Lys Asn Ile Lys Phe Pro Ala Phe Ala Gln Leu Lys Ala 
145                 150                 155                 160 


Asp Gly Ala Arg Cys Phe Ala Glu Val Arg Gly Asp Glu Leu Asp Asp 
                165                 170                 175     


Val Arg Leu Leu Ser Arg Lys Gly Asn Glu Tyr Leu Gly Leu Asp Leu 
            180                 185                 190         


Leu Lys Glu Glu Leu Ile Lys Met Thr Ala Glu Ala Arg Gln Ile His 
        195                 200                 205             


Pro Glu Gly Val Leu Ile Asp Gly Glu Leu Val Tyr His Glu Gln Val 
    210                 215                 220                 


Ala Lys Glu Pro Glu Gly Leu Lys Ala Leu Phe Asp Arg Tyr Pro Glu 
225                 230                 235                 240 


Asn Ser Lys Ala Lys Glu Phe Ala Glu Val Ala Glu Ser Arg Thr Ala 
                245                 250                 255     


Ser Asn Gly Ile Ala Asn Lys Ser Leu Lys Gly Thr Ile Ser Glu Lys 
            260                 265                 270         


Glu Ala Gln Cys Met Lys Phe Gln Val Trp Asp Tyr Val Pro Leu Val 
        275                 280                 285             


Glu Ile Tyr Ser Leu Pro Ala Phe Arg Leu Lys Tyr Asp Val Arg Phe 
    290                 295                 300                 


Ser Lys Leu Glu Gln Met Thr Ser Gly Tyr Asp Lys Val Ile Leu Ile 
305                 310                 315                 320 


Glu Asn Gln Val Val Asn Asn Leu Leu Glu Ala Lys Val Ile Tyr Lys 
                325                 330                 335     


Lys Tyr Ile Asp Gln Gly Leu Glu Gly Ile Ile Leu Lys Asn Ile Asp 
            340                 345                 350         


Gly Leu Trp Glu Asn Ala Arg Ser Lys Asn Leu Tyr Lys Phe Lys Glu 
        355                 360                 365             


Val Ile Trp Val Asp Leu Lys Ile Val Gly Ile Tyr Pro His Arg Lys 
    370                 375                 380                 


Asp Pro Thr Lys Ala Gly Gly Phe Ile Leu Glu Ser Glu Cys Gly Lys 
385                 390                 395                 400 


Ile Lys Val Asn Ala Gly Ser Gly Leu Lys Asp Lys Ala Gly Val Lys 
                405                 410                 415     


Ser His Glu Leu Asp Arg Thr Arg Ile Met Glu Asn Gln Asn Tyr Tyr 
            420                 425                 430         


Ile Gly Lys Ile Leu Glu Cys Lys Cys Asn Gly Trp Leu Lys Ser Asp 
        435                 440                 445             


Gly Arg Lys Pro Gly Val Lys Leu Phe Leu Pro Ile Ala Ile Arg Leu 
    450                 455                 460                 


Arg Gly Asp Lys Thr Lys Ala Asn Thr Phe Glu Asp Val Phe Gly Asp 
465                 470                 475                 480 


Phe His Glu Val Thr Gly Leu 
                485         


