                               SEQUENCE LISTING

<110> UNIVERSITY OF KENTUCKY RESEARCH FOUNDATION
 
<120> PROTEIN VARIANTS FOR USE AS LIPID BILAYER-INTEGRATED NANOPORE, 
      AND METHODS THEREOF

<130> 2935720-000005

<140> PCT/US2016/052158
<141> 2016-09-16

<150> 62/220,545
<151> 2015-09-18

<160> 11    

<170> PatentIn version 3.5

<210> 1
<211> 1643
<212> DNA
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      polynucleotide

<400> 1
gtaacctgca tatggctgat tcaaaacgta caggattggg cgaagacggt gctaaagcta       60

cctatgaccg cctaacaaac gaccgtagag cctatgagac tcgtgcggag aactgtgcgc      120

aatacaccat tccgtccttg ttcccgaagg agtccgataa cgaatctacc gactacacga      180

ctccgtggca ggctgtaggt gcgcggggtc tcaacaatct agcctctaag ttaatgcttg      240

cgttattccc gatgcagtcg tggatgaagc tgaccattag cgaatatgag gcgaagcagc      300

ttgttggaga ccctgatgga ctcgctaagg tggacgaagg tctgtcaatg gttgagcgca      360

taatcatgaa ctatatcgaa tccaacagtt accgcgtaac actctttgag tgcctcaagc      420

agttgatcgt ggctggtaac gccctgcttt acttaccgga accagaaggt agctacaatc      480

cgatgaagct gtaccgattg tcttcttatg ttgtccaaag agacgcatac ggcaatgtgt      540

tacagattgt cactcgtgac cagatagcct ttggtgctct cccggaagac gttaggtctg      600

cggtagagaa atctggtggt gagaagaaga tggacgaaat ggtcgatgtg tacacccatg      660

tgtatctcga tgaagagtcc ggcgattacc tcaagtacga ggaagtagag gacgttgaga      720

ttgatggttc cgatgccacc tatccgactg acgcgatgcc ctacattccg gttcgcatgg      780

ttcgcattga tggcgagtct tacggtcgct cctactgtga agaatactta ggtgacttaa      840

ggtcgcttga gaatctccaa gaggctatcg ttaagatgag tatgattagc gcgaaggtca      900

ttggtctggt gaacccggct ggcattacgc agccccgtag attaaccaaa gctcagactg      960

gtgacttcgt tccaggccgt cgagaagata ttgacttcct gcaactggag aagcaagctg     1020

actttaccgt agcgaaagct gtgagtgacc agatagaagc acgcttatcg tatgccttta     1080

tgttgaactc tgcggtacag cgcacaggcg aacgtgtgac cgccgaagag attcgatacg     1140

ttgcgtcaga actggaagat acgcttggtg gcgtctactc gattctgtct caagaattgc     1200

aattgcctct ggtacgtgtg ctcttgaagc aactccaagc aacctcgcag attcctgagc     1260

taccgaaaga agccgttgag cctactatca gtacaggtct ggaagcaatt ggtcgtggtc     1320

aagacctcga taagctggag cgttgcatct cagcgtgggc ggctcttgcc cctatgcagg     1380

gagacccgga cattaatctt gctgtcatta agctacgcat tgctaacgct ataggtattg     1440

atacttctgg tatcctactg acggatgaac agaagcaagc ccttatgatg caggatgcgg     1500

cacaaacagg cgtcgagaat gctgcggctg ctggtggtgc tggtgttggt gctttggcta     1560

cctcaagtcc agaagccatg caaggtgctg ctgccaaggc tggcctcaac gccaccggtg     1620

gccaccatca ccatcaccat tag                                             1643


<210> 2
<211> 543
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      polypeptide

<400> 2
Met Ala Asp Ser Lys Arg Thr Gly Leu Gly Glu Asp Gly Ala Lys Ala 
1               5                   10                  15      


Thr Tyr Asp Arg Leu Thr Asn Asp Arg Arg Ala Tyr Glu Thr Arg Ala 
            20                  25                  30          


Glu Asn Cys Ala Gln Tyr Thr Ile Pro Ser Leu Phe Pro Lys Glu Ser 
        35                  40                  45              


Asp Asn Glu Ser Thr Asp Tyr Thr Thr Pro Trp Gln Ala Val Gly Ala 
    50                  55                  60                  


Arg Gly Leu Asn Asn Leu Ala Ser Lys Leu Met Leu Ala Leu Phe Pro 
65                  70                  75                  80  


Met Gln Ser Trp Met Lys Leu Thr Ile Ser Glu Tyr Glu Ala Lys Gln 
                85                  90                  95      


Leu Val Gly Asp Pro Asp Gly Leu Ala Lys Val Asp Glu Gly Leu Ser 
            100                 105                 110         


Met Val Glu Arg Ile Ile Met Asn Tyr Ile Glu Ser Asn Ser Tyr Arg 
        115                 120                 125             


Val Thr Leu Phe Glu Cys Leu Lys Gln Leu Ile Val Ala Gly Asn Ala 
    130                 135                 140                 


Leu Leu Tyr Leu Pro Glu Pro Glu Gly Ser Tyr Asn Pro Met Lys Leu 
145                 150                 155                 160 


Tyr Arg Leu Ser Ser Tyr Val Val Gln Arg Asp Ala Tyr Gly Asn Val 
                165                 170                 175     


Leu Gln Ile Val Thr Arg Asp Gln Ile Ala Phe Gly Ala Leu Pro Glu 
            180                 185                 190         


Asp Val Arg Ser Ala Val Glu Lys Ser Gly Gly Glu Lys Lys Met Asp 
        195                 200                 205             


Glu Met Val Asp Val Tyr Thr His Val Tyr Leu Asp Glu Glu Ser Gly 
    210                 215                 220                 


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


Asp Ala Thr Tyr Pro Thr Asp Ala Met Pro Tyr Ile Pro Val Arg Met 
                245                 250                 255     


Val Arg Ile Asp Gly Glu Ser Tyr Gly Arg Ser Tyr Cys Glu Glu Tyr 
            260                 265                 270         


Leu Gly Asp Leu Arg Ser Leu Glu Asn Leu Gln Glu Ala Ile Val Lys 
        275                 280                 285             


Met Ser Met Ile Ser Ala Lys Val Ile Gly Leu Val Asn Pro Ala Gly 
    290                 295                 300                 


Ile Thr Gln Pro Arg Arg Leu Thr Lys Ala Gln Thr Gly Asp Phe Val 
305                 310                 315                 320 


Pro Gly Arg Arg Glu Asp Ile Asp Phe Leu Gln Leu Glu Lys Gln Ala 
                325                 330                 335     


Asp Phe Thr Val Ala Lys Ala Val Ser Asp Gln Ile Glu Ala Arg Leu 
            340                 345                 350         


Ser Tyr Ala Phe Met Leu Asn Ser Ala Val Gln Arg Thr Gly Glu Arg 
        355                 360                 365             


Val Thr Ala Glu Glu Ile Arg Tyr Val Ala Ser Glu Leu Glu Asp Thr 
    370                 375                 380                 


Leu Gly Gly Val Tyr Ser Ile Leu Ser Gln Glu Leu Gln Leu Pro Leu 
385                 390                 395                 400 


Val Arg Val Leu Leu Lys Gln Leu Gln Ala Thr Ser Gln Ile Pro Glu 
                405                 410                 415     


Leu Pro Lys Glu Ala Val Glu Pro Thr Ile Ser Thr Gly Leu Glu Ala 
            420                 425                 430         


Ile Gly Arg Gly Gln Asp Leu Asp Lys Leu Glu Arg Cys Ile Ser Ala 
        435                 440                 445             


Trp Ala Ala Leu Ala Pro Met Gln Gly Asp Pro Asp Ile Asn Leu Ala 
    450                 455                 460                 


Val Ile Lys Leu Arg Ile Ala Asn Ala Ile Gly Ile Asp Thr Ser Gly 
465                 470                 475                 480 


Ile Leu Leu Thr Asp Glu Gln Lys Gln Ala Leu Met Met Gln Asp Ala 
                485                 490                 495     


Ala Gln Thr Gly Val Glu Asn Ala Ala Ala Ala Gly Gly Ala Gly Val 
            500                 505                 510         


Gly Ala Leu Ala Thr Ser Ser Pro Glu Ala Met Gln Gly Ala Ala Ala 
        515                 520                 525             


Lys Ala Gly Leu Asn Ala Thr Gly Gly His His His His His His 
    530                 535                 540             


<210> 3
<211> 1599
<212> DNA
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      polynucleotide

<400> 3
atgaaattta atgtattaag tttgtttgct ccatgggcta aaatggacga acgaaatttt       60

aaagaccaag aaaaagaaga tcttgtttcc attacagccc caaagcttga tgatggagca      120

agagaatttg aagtaagctc gaatgaagct gcttctcctt ataatgctgc attccaaaca      180

atttttggtt catatgaacc aggaatgaaa actactcgtg agcttattga tacatatcgt      240

aatctcatga ataactatga agtagataat gcagtttcag aaatcgtttc agatgctatc      300

gtctatgaag atgatactga agtcgtagcg ttaaatttgg ataaatctaa atttagccca      360

aaaattaaaa atatgatgtt agatgaattt agtgatgtat taaatcatct atcgtttcaa      420

cgaaaaggtt ctgatcattt tagacgttgg tatgttgatt caagaatttt ctttcataaa      480

atcattgatc caaaacgtcc aaaagaaggc ataaaagaat tacgtagatt agaccctcgc      540

caagttcagt atgttcgtga aattataaca gaaactgaag ctggcacaaa aatagttaaa      600

ggttacaaag aatattttat atatgatact gcccatgagt catatgcatg tgatggtaga      660

atgtatgaag ctggcacaaa aataaaaatt cctaaagctg ccgtcgttta tgcccattct      720

ggattagtcg attgttgcgg taaaaatatc atcgggtatt tgcatcgtgc tgttaaacct      780

gctaaccaat taaaattatt agaagatgct gtagtcattt atcgcattac tcgtgctcct      840

gaccgtcgtg tttggtatgt agacacaggt aatatgcctg ctcgtaaagc tgctgagcac      900

atgcaacatg ttatgaacac gatgaaaaac cgtgtagtat atgatgcatc aacaggtaaa      960

ataaaaaatc aacagcataa tatgtctatg accgaagact attggttgca gcgccgtgat     1020

ggtaaagctg tgacagaagt tgatactctt cctggtgctg ataatactgg caatatggaa     1080

gatattcgtt ggtttagaca agctctttat atggcattac gtgttcctct ttcacgcatt     1140

ccgcaagacc aacaaggcgg tgtgatgttt gattctggaa ctagcattac acgtgatgaa     1200

ttaacgtttg ctaaatttat tcgtgagtta cagcacaagt ttgaagaagt tttcctagat     1260

ccgcttaaaa caaatctttt gcttaaaggt ataatcacag aagatgagtg gaatgatgaa     1320

ataaataata ttaagataga atttcatcgg gatagctact ttgctgagct caaagaagca     1380

gaaattttgg aacgaagaat taatatgcta accatggcag aaccatttat tggtaaatat     1440

atttctcaca gaactgctat gaaagacatt ttgcagatga ctgatgaaga aatagaacaa     1500

gaagccaagc aaattgaaga agagtctaaa gaggctcgtt tccaagaccc cgaccaagaa     1560

caagaggatt ttggtggcca ccatcaccat caccattag                            1599


<210> 4
<211> 532
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      polypeptide

<400> 4
Met Lys Phe Asn Val Leu Ser Leu Phe Ala Pro Trp Ala Lys Met Asp 
1               5                   10                  15      


Glu Arg Asn Phe Lys Asp Gln Glu Lys Glu Asp Leu Val Ser Ile Thr 
            20                  25                  30          


Ala Pro Lys Leu Asp Asp Gly Ala Arg Glu Phe Glu Val Ser Ser Asn 
        35                  40                  45              


Glu Ala Ala Ser Pro Tyr Asn Ala Ala Phe Gln Thr Ile Phe Gly Ser 
    50                  55                  60                  


Tyr Glu Pro Gly Met Lys Thr Thr Arg Glu Leu Ile Asp Thr Tyr Arg 
65                  70                  75                  80  


Asn Leu Met Asn Asn Tyr Glu Val Asp Asn Ala Val Ser Glu Ile Val 
                85                  90                  95      


Ser Asp Ala Ile Val Tyr Glu Asp Asp Thr Glu Val Val Ala Leu Asn 
            100                 105                 110         


Leu Asp Lys Ser Lys Phe Ser Pro Lys Ile Lys Asn Met Met Leu Asp 
        115                 120                 125             


Glu Phe Ser Asp Val Leu Asn His Leu Ser Phe Gln Arg Lys Gly Ser 
    130                 135                 140                 


Asp His Phe Arg Arg Trp Tyr Val Asp Ser Arg Ile Phe Phe His Lys 
145                 150                 155                 160 


Ile Ile Asp Pro Lys Arg Pro Lys Glu Gly Ile Lys Glu Leu Arg Arg 
                165                 170                 175     


Leu Asp Pro Arg Gln Val Gln Tyr Val Arg Glu Ile Ile Thr Glu Thr 
            180                 185                 190         


Glu Ala Gly Thr Lys Ile Val Lys Gly Tyr Lys Glu Tyr Phe Ile Tyr 
        195                 200                 205             


Asp Thr Ala His Glu Ser Tyr Ala Cys Asp Gly Arg Met Tyr Glu Ala 
    210                 215                 220                 


Gly Thr Lys Ile Lys Ile Pro Lys Ala Ala Val Val Tyr Ala His Ser 
225                 230                 235                 240 


Gly Leu Val Asp Cys Cys Gly Lys Asn Ile Ile Gly Tyr Leu His Arg 
                245                 250                 255     


Ala Val Lys Pro Ala Asn Gln Leu Lys Leu Leu Glu Asp Ala Val Val 
            260                 265                 270         


Ile Tyr Arg Ile Thr Arg Ala Pro Asp Arg Arg Val Trp Tyr Val Asp 
        275                 280                 285             


Thr Gly Asn Met Pro Ala Arg Lys Ala Ala Glu His Met Gln His Val 
    290                 295                 300                 


Met Asn Thr Met Lys Asn Arg Val Val Tyr Asp Ala Ser Thr Gly Lys 
305                 310                 315                 320 


Ile Lys Asn Gln Gln His Asn Met Ser Met Thr Glu Asp Tyr Trp Leu 
                325                 330                 335     


Gln Arg Arg Asp Gly Lys Ala Val Thr Glu Val Asp Thr Leu Pro Gly 
            340                 345                 350         


Ala Asp Asn Thr Gly Asn Met Glu Asp Ile Arg Trp Phe Arg Gln Ala 
        355                 360                 365             


Leu Tyr Met Ala Leu Arg Val Pro Leu Ser Arg Ile Pro Gln Asp Gln 
    370                 375                 380                 


Gln Gly Gly Val Met Phe Asp Ser Gly Thr Ser Ile Thr Arg Asp Glu 
385                 390                 395                 400 


Leu Thr Phe Ala Lys Phe Ile Arg Glu Leu Gln His Lys Phe Glu Glu 
                405                 410                 415     


Val Phe Leu Asp Pro Leu Lys Thr Asn Leu Leu Leu Lys Gly Ile Ile 
            420                 425                 430         


Thr Glu Asp Glu Trp Asn Asp Glu Ile Asn Asn Ile Lys Ile Glu Phe 
        435                 440                 445             


His Arg Asp Ser Tyr Phe Ala Glu Leu Lys Glu Ala Glu Ile Leu Glu 
    450                 455                 460                 


Arg Arg Ile Asn Met Leu Thr Met Ala Glu Pro Phe Ile Gly Lys Tyr 
465                 470                 475                 480 


Ile Ser His Arg Thr Ala Met Lys Asp Ile Leu Gln Met Thr Asp Glu 
                485                 490                 495     


Glu Ile Glu Gln Glu Ala Lys Gln Ile Glu Glu Glu Ser Lys Glu Ala 
            500                 505                 510         


Arg Phe Gln Asp Pro Asp Gln Glu Gln Glu Asp Phe Gly Gly His His 
        515                 520                 525             


His His His His 
    530         


<210> 5
<211> 9
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      peptide

<400> 5
Trp Ser His Pro Gln Arg Phe Glu Lys 
1               5                   


<210> 6
<211> 12
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      peptide


<220>
<221> misc_feature
<222> (1)..(12) 
<223> This sequence may encompass 3-12 contiguous histidine
      residues 

<400> 6
His His His His His His His His His His His His 
1               5                   10          


<210> 7
<211> 12
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      peptide


<220>
<221> misc_feature
<222> (1)..(12) 
<223> This sequence may encompass 3-12 contiguous arginine 
      residues 

<400> 7
Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg 
1               5                   10          


<210> 8
<211> 10
<212> PRT
<213> Human Immunodeficiency virus 

<400> 8
Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg 
1               5                   10  


<210> 9
<211> 6
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      peptide

<400> 9
Asp Arg Ala Thr Pro Tyr 
1               5       


<210> 10
<211> 6
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      6xHis tag

<400> 10
His His His His His His 
1               5       


<210> 11
<211> 6
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      peptide

<400> 11
Gly Gly Gly Gly Gly Gly 
1               5       


