                         SEQUENCE LISTING

<110>  Agency for Science Technology and Research
       Nanyang Technological University
 
<120>  Mutants of The Bacteriophage Lambda Integrase

<130>  9869SG2408

<160>  31    

<170>  PatentIn version 3.5

<210>  1
<211>  356
<212>  PRT
<213>  Bacteriophage lambda (Integrase-h/218)

<400>  1

Met Gly Arg Arg Arg Ser His Glu Arg Arg Asp Leu Pro Pro Asn Leu 
1               5                   10                  15      


Tyr Ile Arg Asn Asn Gly Tyr Tyr Cys Tyr Arg Asp Pro Arg Thr Gly 
            20                  25                  30          


Lys Glu Phe Gly Leu Gly Arg Asp Arg Arg Ile Ala Ile Thr Glu Ala 
        35                  40                  45              


Ile Gln Ala Asn Ile Glu Leu Phe Ser Gly His Lys His Lys Pro Leu 
    50                  55                  60                  


Thr Ala Arg Ile Asn Ser Asp Asn Ser Val Thr Leu His Ser Trp Leu 
65                  70                  75                  80  


Asp Arg Tyr Glu Lys Ile Leu Ala Ser Arg Gly Ile Lys Gln Lys Thr 
                85                  90                  95      


Leu Ile Asn Tyr Met Ser Lys Ile Lys Ala Ile Arg Arg Gly Leu Pro 
            100                 105                 110         


Asp Ala Pro Leu Glu Asp Ile Thr Thr Lys Glu Ile Ala Ala Met Leu 
        115                 120                 125             


Asn Gly Tyr Ile Asp Glu Gly Lys Ala Ala Ser Ala Lys Leu Ile Arg 
    130                 135                 140                 


Ser Thr Leu Ser Asp Ala Phe Arg Glu Ala Ile Ala Glu Gly His Ile 
145                 150                 155                 160 


Thr Thr Asn His Val Ala Ala Thr Arg Ala Ala Lys Ser Lys Val Arg 
                165                 170                 175     


Arg Ser Arg Leu Thr Ala Asp Glu Tyr Leu Lys Ile Tyr Gln Ala Ala 
            180                 185                 190         


Glu Ser Ser Pro Cys Trp Leu Arg Leu Ala Met Glu Leu Ala Val Val 
        195                 200                 205             


Thr Gly Gln Arg Val Gly Asp Leu Cys Lys Met Lys Trp Ser Asp Ile 
    210                 215                 220                 


Val Asp Gly Tyr Leu Tyr Val Glu Gln Ser Lys Thr Gly Val Lys Ile 
225                 230                 235                 240 


Ala Ile Pro Thr Ala Leu His Ile Asp Ala Leu Gly Ile Ser Met Lys 
                245                 250                 255     


Glu Thr Leu Asp Lys Cys Lys Glu Ile Leu Gly Gly Glu Thr Ile Ile 
            260                 265                 270         


Ala Ser Thr Arg Arg Glu Pro Leu Ser Ser Gly Thr Val Ser Arg Tyr 
        275                 280                 285             


Phe Met Arg Ala Arg Lys Ala Ser Gly Leu Ser Phe Glu Gly Asp Pro 
    290                 295                 300                 


Pro Thr Phe His Glu Leu Arg Ser Leu Ser Ala Arg Leu Tyr Glu Lys 
305                 310                 315                 320 


Gln Ile Ser Asp Lys Phe Ala Gln His Leu Leu Gly His Lys Ser Asp 
                325                 330                 335     


Thr Met Ala Ser Gln Tyr Arg Asp Asp Arg Gly Arg Glu Trp Asp Lys 
            340                 345                 350         


Ile Glu Ile Lys 
        355     


<210>  2
<211>  356
<212>  PRT
<213>  artificial

<220>
<223>  C2 lambda integrase mutant

<400>  2

Met Gly Arg Arg Arg Ser His Glu Arg Arg Asp Leu Pro Pro Asn Leu 
1               5                   10                  15      


Tyr Ile Arg Asn Asn Gly Tyr Tyr Cys Tyr Arg Asp Pro Arg Thr Gly 
            20                  25                  30          


Lys Glu Phe Gly Leu Gly Arg Asp Arg Arg Ile Ala Ile Thr Glu Ala 
        35                  40                  45              


Ile Gln Ala Asn Ile Glu Leu Phe Ser Gly His Lys His Lys Pro Leu 
    50                  55                  60                  


Thr Ala Arg Ile Asn Ser Asp Asn Ser Val Thr Leu His Ser Trp Leu 
65                  70                  75                  80  


Asp Arg Tyr Glu Lys Ile Leu Ala Ser Arg Gly Ile Lys Gln Lys Thr 
                85                  90                  95      


Leu Ile Asn Tyr Met Ser Lys Ile Lys Ala Ile Arg Arg Gly Leu Pro 
            100                 105                 110         


Asp Ala Pro Leu Glu Asp Ile Thr Thr Lys Glu Ile Ala Ala Met Leu 
        115                 120                 125             


Asn Gly Tyr Ile Asp Glu Gly Lys Ala Ala Ser Ala Lys Leu Ile Arg 
    130                 135                 140                 


Ser Thr Leu Ser Asp Ala Phe Arg Glu Ala Ile Ala Glu Gly His Ile 
145                 150                 155                 160 


Thr Thr Asn His Val Ala Ala Thr Arg Ala Ala Lys Ser Lys Val Arg 
                165                 170                 175     


Arg Ser Arg Leu Thr Ala Asp Glu Tyr Leu Lys Ile Tyr Gln Ala Ala 
            180                 185                 190         


Glu Ser Ser Pro Cys Trp Leu Arg Leu Ala Met Glu Leu Ala Val Val 
        195                 200                 205             


Thr Gly Gln Arg Val Gly Asp Leu Cys Lys Met Lys Trp Ser Asp Ile 
    210                 215                 220                 


Val Asp Gly Tyr Leu Tyr Val Glu Gln Ser Lys Thr Gly Val Lys Ile 
225                 230                 235                 240 


Ala Ile Pro Thr Ala Leu His Ile Asp Ala Leu Gly Ile Ser Met Lys 
                245                 250                 255     


Glu Thr Leu Asp Lys Cys Lys Glu Ile Leu Gly Gly Glu Thr Ile Ile 
            260                 265                 270         


Ala Ser Thr Arg Arg Glu Pro Leu Ser Ser Gly Thr Val Ser Arg Tyr 
        275                 280                 285             


Phe Met Arg Ala Arg Lys Ala Ser Gly Leu Ser Phe Glu Gly Asp Pro 
    290                 295                 300                 


Pro Thr Phe His Glu Leu Arg Ser Leu Ser Ala Arg Leu Tyr Glu Lys 
305                 310                 315                 320 


Gln Ile Ser Asp Lys Phe Ala Gln His Leu Leu Gly His Lys Ser Val 
                325                 330                 335     


Thr Met Ala Ser Gln Tyr Arg Asp Asp Arg Gly Arg Glu Trp Asp Lys 
            340                 345                 350         


Ile Glu Ile Lys 
        355     


<210>  3
<211>  356
<212>  PRT
<213>  artificial

<220>
<223>  C3 lambda integrase mutant

<400>  3

Met Gly Arg Arg Arg Ser His Glu Arg Arg Asp Leu Pro Pro Asn Leu 
1               5                   10                  15      


Tyr Ile Arg Asn Asn Gly Tyr Tyr Cys Tyr Arg Asp Pro Arg Thr Gly 
            20                  25                  30          


Lys Glu Phe Gly Leu Gly Arg Asp Arg Arg Phe Ala Ile Thr Glu Ala 
        35                  40                  45              


Ile Gln Ala Asn Ile Glu Leu Phe Ser Gly His Lys His Lys Pro Leu 
    50                  55                  60                  


Thr Ala Arg Ile Asn Ser Asp Asn Ser Val Thr Leu His Ser Trp Leu 
65                  70                  75                  80  


Asp Arg Tyr Glu Lys Ile Leu Ala Ser Arg Gly Ile Lys Gln Lys Thr 
                85                  90                  95      


Leu Ile Asn Tyr Met Ser Lys Ile Lys Ala Ile Arg Arg Gly Leu Pro 
            100                 105                 110         


Asp Ala Pro Leu Glu Asp Ile Thr Thr Lys Glu Ile Ala Ala Met Leu 
        115                 120                 125             


Asn Gly Tyr Ile Asp Glu Gly Lys Ala Ala Ser Ala Lys Leu Ile Arg 
    130                 135                 140                 


Ser Thr Leu Ser Asp Ala Phe Arg Glu Ala Ile Ala Glu Gly His Ile 
145                 150                 155                 160 


Thr Thr Asn His Val Ala Ala Thr Arg Ala Ala Lys Ser Lys Val Arg 
                165                 170                 175     


Arg Ser Arg Leu Thr Ala Asp Glu Tyr Leu Lys Ile Tyr Gln Ala Ala 
            180                 185                 190         


Glu Ser Ser Pro Cys Trp Leu Arg Leu Ala Met Glu Leu Ala Val Val 
        195                 200                 205             


Thr Gly Gln Arg Val Gly Asp Leu Cys Lys Met Lys Trp Ser Asp Ile 
    210                 215                 220                 


Val Asp Gly Tyr Leu Tyr Val Glu Gln Ser Lys Thr Gly Val Lys Ile 
225                 230                 235                 240 


Ala Ile Pro Thr Ala Leu His Ile Asp Ala Leu Gly Ile Ser Met Lys 
                245                 250                 255     


Glu Thr Leu Asp Lys Cys Lys Glu Ile Leu Gly Gly Glu Thr Ile Ile 
            260                 265                 270         


Ala Ser Thr Arg Arg Glu Pro Leu Ser Ser Gly Thr Val Ser Arg Tyr 
        275                 280                 285             


Phe Met Arg Ala Arg Lys Ala Ser Gly Leu Ser Phe Glu Gly Asp Pro 
    290                 295                 300                 


Pro Thr Phe His Glu Leu Arg Ser Leu Ser Ala Arg Leu Tyr Gly Lys 
305                 310                 315                 320 


Gln Ile Ser Asp Lys Phe Ala Gln His Leu Leu Gly His Lys Ser Val 
                325                 330                 335     


Thr Met Ala Ser Gln Tyr Arg Asp Asp Arg Gly Arg Glu Trp Asp Lys 
            340                 345                 350         


Ile Glu Ile Lys 
        355     


<210>  4
<211>  1071
<212>  DNA
<213>  Bacteriophage lambda integrase

<400>  4
atgggaagaa ggcgaagtca tgagcgccgg gatttacccc ctaaccttta tataagaaac       60

aatggatatt actgctacag ggacccaagg acgggtaaag agtttggatt aggcagagac      120

aggcgaatcg caatcactga agctatacag gccaacattg agttattttc aggacacaaa      180

cacaagcctc tgacagcgag aatcaacagt gataattccg ttacgttaca ttcatggctt      240

gatcgctacg aaaaaatcct ggccagcaga ggaatcaagc agaagacact cataaattac      300

atgagcaaaa ttaaagcaat aaggaggggt ctgcctgatg ctccacttga agacatcacc      360

acaaaagaaa ttgcggcaat gctcaatgga tacatagacg agggcaaggc ggcgtcagcc      420

aagttaatca gatcaacact gagcgatgca ttccgagagg caatagctga aggccatata      480

acaacaaacc atgtcgctgc cactcgcgca gcaaaatcaa aggtaaggag atcaagactt      540

acggctgacg aatacctgaa aatttatcaa gcagcagaat catcaccatg ttggctcaga      600

cttgcaatgg aactggctgt tgttaccggg caacgagttg gtgatttatg caaaatgaag      660

tggtctgata tcgtagatgg atatctttat gtcgagcaaa gcaaaacagg cgtaaaaatt      720

gccatcccaa cagcattgca tattgatgct ctcggaatat caatgaagga aacacttgat      780

aaatgcaaag agattcttgg cggagaaacc ataattgcat ctactcgtcg cgaaccgctt      840

tcatccggca cagtatcaag gtattttatg cgcgcacgaa aagcatcagg tctttccttc      900

gaaggggatc cgcctacctt tcacgagttg cgcagtttgt ctgcaagact ctatgagaag      960

cagataagcg ataagtttgc tcaacatctt ctcgggcata agtcggacac catggcatca     1020

cagtatcgtg atgacagagg cagggagtgg gacaaaattg aaatcaaata a              1071


<210>  5
<211>  21
<212>  DNA
<213>  attB

<400>  5
ctgctttttt atactaactt g                                                 21


<210>  6
<211>  21
<212>  DNA
<213>  attP

<400>  6
cagctttttt atactaagtt g                                                 21


<210>  7
<211>  21
<212>  DNA
<213>  attH

<400>  7
ctgctttctt ataccaagtg g                                                 21


<210>  8
<211>  21
<212>  DNA
<213>  attPH

<400>  8
cagctttctt ataccaagtt g                                                 21


<210>  9
<211>  21
<212>  DNA
<213>  attP4X

<400>  9
cagctttatt tcattaagtt g                                                 21


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

<220>
<223>  petF2 primer

<400>  10
catcggtgat gtcggcgat                                                    19


<210>  11
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  petR primer

<400>  11
cggatatagt tcctcctttc agca                                              24


<210>  12
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  attP-F primer

<400>  12
cacagaattc cgtctgttac aggtcactaa taccatct                               38


<210>  13
<211>  52
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  attPSOE-R primer

<400>  13
acatttcccc gaaaagtgcc acctgaacat caccgggaaa tcaaataatg at               52


<210>  14
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TEM1prom-F primer

<400>  14
ttcaggtggc acttttcggg gaaatgt                                           27


<210>  15
<211>  41
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TEM1prom-R primer

<400>  15
tgtggaattc ctacactaga aggacagtat ttggtatctg c                           41


<210>  16
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EcoliAttB-F primer

<400>  16
ctgaaaatgt gttcacaggt tgct                                              24


<210>  17
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EcoliAttB-R primer

<400>  17
gcaatgccat ctggtatcac t                                                 21


<210>  18
<211>  1071
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  C2 mutant gene

<400>  18
atgggaagaa ggcgaagtca tgagcgccgg gatttacccc ctaaccttta tataagaaac       60

aatggatatt actgctacag ggacccaagg acgggtaaag agtttggatt aggcagagac      120

aggcgaatcg caatcactga agctatacag gccaacattg agttattttc aggacacaaa      180

cacaagcctc tgacagcgag aatcaacagt gataattccg ttacgttaca ttcatggctt      240

gatcgctacg aaaaaatcct ggccagcaga ggaatcaagc agaagacact cataaattac      300

atgagcaaaa ttaaagcaat aaggaggggt ctgcctgatg ctccacttga agacatcacc      360

acaaaagaaa ttgcggcaat gctcaatgga tacatagacg agggcaaggc ggcgtcagcc      420

aagttaatca gatcaacgct gagcgatgca ttccgagagg caatagctga aggccatata      480

acaacaaacc atgtcgctgc cactcgcgca gcaaagtcaa aggtaaggag atcaagactt      540

acggctgacg aatacctgaa aatttatcaa gcagcagaat catcaccatg ttggctcaga      600

cttgcaatgg aactggctgt tgttaccggg caacgagttg gtgacttgtg caaaatgaag      660

tggtctgata tcgtagatgg atatctttat gtcgagcaaa gcaaaacagg cgtaaaaatt      720

gccatcccaa cagcattgca tattgatgct ctcggaatat caatgaagga aacacttgat      780

aaatgcaaag agattcttgg cggagaaacc ataattgcat ctactcgtcg cgaaccgctt      840

tcatccggca cagtatcaag gtattttatg cgcgcacgaa aagcatcagg tctttccttc      900

gaaggggatc cgcctacctt tcacgagttg cgcagtttgt ctgcaagact ctatgagaag      960

cagataagcg ataagtttgc tcaacatctt ctcgggcata agtcggtcac catggcatca     1020

cagtatcgtg atgacagagg cagggagtgg gacaaaattg aaatcaaata a              1071


<210>  19
<211>  1071
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  C3 mutant gene

<400>  19
atgggaagaa ggcgaagtca tgagcgccgg gatttacccc ctaaccttta tataagaaac       60

aatggatatt actgctacag ggacccaagg acgggtaaag agtttggatt aggcagagac      120

aggcgattcg caatcactga agctatacag gccaacattg agttattttc aggacacaaa      180

cacaagcctc tgacagcgag aatcaacagt gataattccg ttacgttaca ttcatggctt      240

gatcgctacg aaaaaatcct ggccagcaga ggaatcaagc agaagacact cataaattac      300

atgagcaaaa ttaaagcaat aaggaggggt ctgcctgatg ctccacttga agacatcacc      360

acaaaagaaa ttgcggcaat gctcaatgga tacatagacg agggcaaggc ggcgtcagcc      420

aagttaatca gatcaacgct gagcgatgca ttccgagagg caatagctga aggccatata      480

acaacaaacc atgtcgctgc cactcgcgcg gcaaagtcaa aggtaaggag atcaagactt      540

acggctgacg aatacctgaa aatttatcaa gcagcagaat catcaccatg ttggctcaga      600

cttgcaatgg aactggctgt tgttaccggg caacgagttg gtgacttgtg caaaatgaag      660

tggtctgata tcgtagatgg atatctttat gtcgagcaaa gcaaaacagg cgtaaaaatt      720

gccatcccaa cagcattgca tattgatgct ctcggaatat caatgaagga aacacttgat      780

aaatgcaaag agattcttgg cggagaaacc ataattgcat ctactcgtcg cgaaccgctc      840

tcatccggca cagtatcaag gtattttatg cgcgcacgaa aagcatcagg tctttccttc      900

gaaggggatc cgcctacctt tcacgagttg cgcagtttgt ctgcaagact ctatgggaag      960

cagataagcg ataagtttgc tcaacatctt ctcgggcata agtcggtcac catggcatca     1020

cagtatcgtg atgacagagg cagggagtgg gacaaaattg aaatcaaata a              1071


<210>  20
<211>  1481
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  C3 mutant minicircle DNA

<400>  20
catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg       60

ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac      120

tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa      180

gaaggagata tacatatggg aagaaggcga agtcatgagc gccgggattt accccctaac      240

ctttatataa gaaacaatgg atattactgc tacagggacc caaggacggg taaagagttt      300

ggattaggca gagacaggcg attcgcaatc actgaagcta tacaggccaa cattgagtta      360

ttttcaggac acaaacacaa gcctctgaca gcgagaatca acagtgataa ttccgttacg      420

ttacattcat ggcttgatcg ctacgaaaaa atcctggcca gcagaggaat caagcagaag      480

acactcataa attacatgag caaaattaaa gcaataagga ggggtctgcc tgatgctcca      540

cttgaagaca tcaccacaaa agaaattgcg gcaatgctca atggatacat agacgagggc      600

aaggcggcgt cagccaagtt aatcagatca acgctgagcg atgcattccg agaggcaata      660

gctgaaggcc atataacaac aaaccatgtc gctgccactc gcgcggcaaa gtcaaaggta      720

aggagatcaa gacttacggc tgacgaatac ctgaaaattt atcaagcagc agaatcatca      780

ccatgttggc tcagacttgc aatggaactg gctgttgtta ccgggcaacg agttggtgac      840

ttgtgcaaaa tgaagtggtc tgatatcgta gatggatatc tttatgtcga gcaaagcaaa      900

acaggcgtaa aaattgccat cccaacagca ttgcatattg atgctctcgg aatatcaatg      960

aaggaaacac ttgataaatg caaagagatt cttggcggag aaaccataat tgcatctact     1020

cgtcgcgaac cgctctcatc cggcacagta tcaaggtatt ttatgcgcgc acgaaaagca     1080

tcaggtcttt ccttcgaagg ggatccgcct acctttcacg agttgcgcag tttgtctgca     1140

agactctatg ggaagcagat aagcgataag tttgctcaac atcttctcgg gcataagtcg     1200

gtcaccatgg catcacagta tcgtgatgac agaggcaggg agtgggacaa aattgaaatc     1260

aaacatcatc accatcacca ctaatgagaa ttcgagctcc gtcgacaagc ttgcggccgc     1320

actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc     1380

tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg     1440

ggtcttgagg ggttttttgc tgaaaggagg aactatatcc g                         1481


<210>  21
<211>  1599
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  attP-TEM1 minicircle

<400>  21
cacagaattc cgtctgttac aggtcactaa taccatctaa gtagttgatt catagtgact       60

gcatatattg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata      120

tattgatatt tatatcattt tacgtttctc gttcagcttt tttatactaa gttggcatta      180

taaaaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa      240

tcattatttg atttcccggt gatgttcagg tggcactttt cggggaaatg tgcgcggaac      300

ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc      360

ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt      420

cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct      480

ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga      540

tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag      600

cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca      660

actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga      720

aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag      780

tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc      840

ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa      900

tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt      960

gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg     1020

gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt     1080

tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg     1140

gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat     1200

ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact     1260

gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa     1320

aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt     1380

ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt     1440

ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg     1500

tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca     1560

gataccaaat actgtccttc tagtgtagcc gtagttagg                            1599


<210>  22
<211>  21
<212>  DNA
<213>  HOP'

<400>  22
atgctttatt tcattaagtt g                                                 21


<210>  23
<211>  103
<212>  DNA
<213>  attL

<400>  23
gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa       60

atacaatcat tatttgattt caattttgtc ccactccctc ccg                        103


<210>  24
<211>  331
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PGK promoter

<400>  24
aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag       60

ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca      120

ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc      180

ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg      240

aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg      300

taggcctttg gggcagcggc caatagcagc t                                     331


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

<220>
<223>  HOP'attH4X_F1 primer

<400>  25
gagtgttttc caacttggtt ccatt                                             25


<210>  26
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PuroRev24 primer

<400>  26
caccgtgggc ttgtactcgg tc                                                22


<210>  27
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pLIR-F1 primer

<400>  27
ctgcatcgat tcagctagct g                                                 21


<210>  28
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pLIR-R1 primer

<400>  28
ctgatagtga cctgttcgtt gc                                                22


<210>  29
<211>  4906
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pPGKssPuro-attP4X (targeting vector)

<400>  29
gaattcctct gttacaggtc actaatacca tctaagtagt tgattcatag tgactgcata       60

tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg      120

atatttatat cattttacgt ttctcgttca gctttatttc attaagttgg cattataaaa      180

aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt      240

atttgatttc aattttgtcc cactccctcc cgaattctac cgggtagggg aggcgctttt      300

cccaaggcag tctggagcat gcgctttagc agccccgctg gcacttggcg ctacacaagt      360

ggcctctggc ctcgcacaca ttccacatcc accggtagcg ccaaccggct ccgttctttg      420

gtggcccctt cgcgccactt ctactcctcc cctagtcagg aagtttcccc cccgccccgc      480

agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga      540

tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc      600

tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca      660

ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca      720

cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga      780

ccaattcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctca aaagcgggca      840

tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata ttcacctggc      900

ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc agaaaagaca atctttttgt      960

tgtcaagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg acaatgacat     1020

ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcaga tgaccgagta     1080

caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc     1140

cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga     1200

gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt     1260

gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc     1320

gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc     1380

cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt     1440

cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt     1500

gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc     1560

gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt     1620

gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat ctagagctcg     1680

ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt     1740

gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat     1800

tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag     1860

caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc     1920

ttctgaggcg gaaagaacca gctggggctc gagatccact agttctagcc tcgaggctag     1980

agcggccgcc accgcggtgg agctccaatt cgccctatag tgagtcgtat tacgcgcgct     2040

cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc     2100

gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc     2160

gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat     2220

taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag     2280

cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc     2340

aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc     2400

ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt     2460

ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa     2520

caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg     2580

cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat     2640

taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt     2700

atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct     2760

tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc     2820

cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa     2880

agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg     2940

taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt     3000

tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg     3060

catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac     3120

ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc     3180

ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa     3240

catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc     3300

aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt     3360

aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga     3420

taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa     3480

atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa     3540

gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa     3600

tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt     3660

ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt     3720

gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg     3780

agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt     3840

aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca     3900

agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac     3960

tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac     4020

atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct     4080

taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg     4140

gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca     4200

gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt     4260

aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta     4320

tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc     4380

gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc     4440

cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa     4500

ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag     4560

cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg     4620

ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga     4680

gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat     4740

gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag     4800

ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctggg     4860

taccgggccc cccctcgagg tcgacggtat cgataagctt gatatc                    4906


<210>  30
<211>  5308
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pCMVssKZ-IntC3-CNLS (integrase expression plasmid)

<400>  30
gaattcctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg       60

ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg      120

gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc      180

cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat      240

tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat      300

catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat      360

gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc      420

gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac      480

tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa      540

aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt      600

aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc      660

tagcaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg      720

catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg      780

gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt      840

gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac      900

atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca gctcgaggtc      960

caccatggga agaaggcgaa gtcatgagcg ccgggattta ccccctaacc tttatataag     1020

aaacaatgga tattactgct acagggaccc aaggacgggt aaagagtttg gattaggcag     1080

agacaggcga ttcgcaatca ctgaagctat acaggccaac attgagttat tttcaggaca     1140

caaacacaag cctctgacag cgagaatcaa cagtgataat tccgttacgt tacattcatg     1200

gcttgatcgc tacgaaaaaa tcctggccag cagaggaatc aagcagaaga cactcataaa     1260

ttacatgagc aaaattaaag caataaggag gggtctgcct gatgctccac ttgaagacat     1320

caccacaaaa gaaattgcgg caatgctcaa tggatacata gacgagggca aggcggcgtc     1380

agccaagtta atcagatcaa cgctgagcga tgcattccga gaggcaatag ctgaaggcca     1440

tataacaaca aaccatgtcg ctgccactcg cgcggcaaag tcaaaggtaa ggagatcaag     1500

acttacggct gacgaatacc tgaaaattta tcaagcagca gaatcatcac catgttggct     1560

cagacttgca atggaactgg ctgttgttac cgggcaacga gttggtgact tgtgcaaaat     1620

gaagtggtct gatatcgtag atggatatct ttatgtcgag caaagcaaaa caggcgtaaa     1680

aattgccatc ccaacagcat tgcatattga tgctctcgga atatcaatga aggaaacact     1740

tgataaatgc aaagagattc ttggcggaga aaccataatt gcatctactc gtcgcgaacc     1800

gctctcatcc ggcacagtat caaggtattt tatgcgcgca cgaaaagcat caggtctttc     1860

cttcgaaggg gatccgccta cctttcacga gttgcgcagt ttgtctgcaa gactctatgg     1920

gaagcagata agcgataagt ttgctcaaca tcttctcggg cataagtcgg tcaccatggc     1980

atcacagtat cgtgatgaca gaggcaggga gtgggacaaa attgaaatca aatccggagg     2040

cggccctaag aagaagagaa aggtatgata atctagagct cgctgatcag cctcgactgt     2100

gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga     2160

aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag     2220

taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga     2280

agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac     2340

cagctggggc tcgagatcca ctagttctag cctcgaggct agagcggccg ccaccgcggt     2400

ggagctccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac     2460

aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc     2520

ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc     2580

gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg     2640

tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt     2700

tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc     2760

tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg     2820

gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg     2880

agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct     2940

cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg     3000

agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg     3060

tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc     3120

aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag     3180

gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg     3240

ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt     3300

gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt     3360

tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt     3420

attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa     3480

tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag     3540

agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac     3600

aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac     3660

tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac     3720

cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac     3780

tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact     3840

tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg     3900

tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt     3960

tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat     4020

aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta     4080

gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa     4140

tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga     4200

aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac     4260

aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt     4320

tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc     4380

gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat     4440

cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag     4500

acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc     4560

cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag     4620

cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac     4680

aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg     4740

gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct     4800

atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc     4860

tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga     4920

gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga     4980

agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg     5040

cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt     5100

gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt     5160

gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc     5220

caagcgcgca attaaccctc actaaaggga acaaaagctg ggtaccgggc cccccctcga     5280

ggtcgacggt atcgataagc ttgatatc                                        5308


<210>  31
<211>  21
<212>  DNA
<213>  attH4X

<400>  31
acgctttatt tcattaagtt g                                                 21


