                         SEQUENCE LISTING

<110>  Nanyang Technological University
 
<120>  METHOD FOR THE PRODUCTION OF SEAMLESS DNA VECTORS

<130>  P122248

<150>  SG10202102572X
<151>  2021-03-12

<160>  22    

<170>  PatentIn version 3.5

<210>  1
<211>  2713
<212>  DNA
<213>  Artificial

<220>
<223>  multi-transgene DNA sequence ISC

<400>  1
ggaagaaggc gaagtcatga gcgccgggat ttacccccta acctttatat aagaaacaat       60

ggatattact gctacaggga cccaaggacg ggtaaagagt ttggattagg cagagacagg      120

cgattcgcaa tcactgaagc tatacaggcc aacattgagt tattttcagg acacaaacac      180

aagcctctga cagcgagaat caacagtgat aattccgtta cgttacattc atggcttgat      240

cgctacgaaa aaatcctggc cagcagagga atcaagcaga agacactcat aaattacatg      300

agcaaaatta aagcaataag gaggggtctg cctgatgctc cacttgaaga catcaccaca      360

aaagaaattg cggcaatgct caatggatac atagacgagg gcaaggcggc gtcagccaag      420

ttaatcagat caacgctgag cgatgcattc cgagaggcaa tagctgaagg ccatataaca      480

acaaaccatg tcgctgccac tcgcgcggca aagtcaaagg taaggagatc aagacttacg      540

gctgacgaat acctgaaaat ttatcaagca gcagaatcat caccatgttg gctcagactt      600

gcaatggaac tggctgttgt taccgggcaa cgagttggtg acttgtgcaa aatgaagtgg      660

tctgatatcg tagatggata tctttatgtc gagcaaagca aaacaggcgt aaaaattgcc      720

atcccaacag cattgcatat tgatgctctc ggaatatcaa tgaaggaaac acttgataaa      780

tgcaaagaga ttcttggcgg agaaaccata attgcatcta ctcgtcgcga accgctctca      840

tccggcacag tatcaaggta ttttatgcgc gcacgaaaag catcaggtct ttccttcgaa      900

ggggatccgc ctacctttca cgagttgcgc agtttgtctg caagactcta tgggaagcag      960

ataagcgata agtttgctca acatcttctc gggcataagt cggtcaccat ggcatcacag     1020

tatcgtgatg acagaggcag ggagtgggac aaaattgaaa tcaaatgaaa aacgaatggc     1080

tagcaccaag tcagaattga tagaaagact tgccacccag caatcgcaca ttcccgccaa     1140

gacggttgaa gatgcagtaa aagagatgct ggagcatatg gcctcgactc ttgcgcaggg     1200

tggaagcggc ggtcttacaa aagctgaaat gtcagaatat ctgtttgata agcttgggct     1260

tagcaagcgg gatgccaaag aactggttga actgtttttc gaagagatcc gtcgcgctct     1320

ggaaaacggc gaacaggtga aactctctgg ttttggtaac ttcgatctgc gtgataagaa     1380

tcaacgcccg ggacgtaacc cgaaaacggg cgaggatatt cccattacag cacggcgcgt     1440

ggtgaccttc agacccgggc agaagttaaa aagccgggtc gaaaacgctg gtgggggcga     1500

gcgtattgaa atccgcggtt tcggcagttt ctctttgcac taccgcgcac cacgtaccgg     1560

acgtaatccg aagactggcg ataaagtaga actggaagga aaatacgttc ctcactttaa     1620

acctggtaaa gaactgcgcg atcgcgccaa tatctacggt ggatcaggct gataaataaa     1680

acgaaaggct cagtcgaaag actgggcctt tcgttttatc ctattccgaa gttcctattc     1740

tctagaaagt ataggaactt cggcgcgcct acctgtgacg gaagatcact tcgcagaata     1800

aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg     1860

agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc     1920

gggcgtattt tttgagttgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa     1980

aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc     2040

atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt     2100

tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc     2160

ccgcctgatg aatgctcatc cggaattacg tatggcaatg aaagacggtg agctggtgat     2220

atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc     2280

gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt     2340

ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt     2400

cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga     2460

caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct     2520

gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat     2580

gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaag gcgcgccatt     2640

taaatgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcgactgta     2700

taaaaccaca gcc                                                        2713


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

<220>
<223>  mutant INTC3 recombinase

<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 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>  3
<211>  72
<212>  DNA
<213>  Artificial

<220>
<223>  Primer sequence

<400>  3
actctctact gtttctccat acccgttttt ttggatggag tgaaacgatg ggaagaaggc       60

gaagtcatga gc                                                           72


<210>  4
<211>  72
<212>  DNA
<213>  Artificial

<220>
<223>  Primer sequence

<400>  4
gccaaagctc gcacagaatc actgccaaaa tcgaggccaa ttgcaatcgc ttatacagtc       60

gaagttccta ta                                                           72


<210>  5
<211>  25
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  5
gtctataatc acggcagaaa agtcc                                             25


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

<220>
<223>  PCR primer

<400>  6
tcgcctgtct ctgcctaatc c                                                 21


<210>  7
<211>  19
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  7
cgcaaggcga caaggtgct                                                    19


<210>  8
<211>  25
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  8
ccgcttccat tgactcaatg tagtc                                             25


<210>  9
<211>  198
<212>  PRT
<213>  Artificial

<220>
<223>  scIHF2

<400>  9

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


Ser His Ile Pro Ala Lys Thr Val Glu Asp Ala Val Lys Glu Met Leu 
            20                  25                  30          


Glu His Met Ala Ser Thr Leu Ala Gln Gly Gly Ser Gly Gly Leu Thr 
        35                  40                  45              


Lys Ala Glu Met Ser Glu Tyr Leu Phe Asp Lys Leu Gly Leu Ser Lys 
    50                  55                  60                  


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


Ala Leu Glu Asn Gly Glu Gln Val Lys Leu Ser Gly Phe Gly Asn Phe 
                85                  90                  95      


Asp Leu Arg Asp Lys Asn Gln Arg Pro Gly Arg Asn Pro Lys Thr Gly 
            100                 105                 110         


Glu Asp Ile Pro Ile Thr Ala Arg Arg Val Val Thr Phe Arg Pro Gly 
        115                 120                 125             


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


Glu Ile Arg Gly Phe Gly Ser Phe Ser Leu His Tyr Arg Ala Pro Arg 
145                 150                 155                 160 


Thr Gly Arg Asn Pro Lys Thr Gly Asp Lys Val Glu Leu Glu Gly Lys 
                165                 170                 175     


Tyr Val Pro His Phe Lys Pro Gly Lys Glu Leu Arg Asp Arg Ala Asn 
            180                 185                 190         


Ile Tyr Gly Gly Ser Gly 
        195             


<210>  10
<211>  1071
<212>  DNA
<213>  Artificial

<220>
<223>  IntC3 coding sequence

<400>  10
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>  11
<211>  261
<212>  DNA
<213>  Artificial

<220>
<223>  attP recombination site

<400>  11
tctgttacag gtcactaata ccatctaagt agttgattca tagtgactgc atatcttgtg       60

ttttacagta ttaattagtc tgttttttat ccaaaatcta atttaatata ttgatattta      120

tatcatttta cgtttctcgt tcagcttttt tatactaagt tggcattata aaaaagcatt      180

gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaatacaatc attatttgat      240

ttcaattttg tcccactccc t                                                261


<210>  12
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  attB recombination site

<400>  12
ctgctttttt atactaactt g                                                 21


<210>  13
<211>  120
<212>  DNA
<213>  Artificial

<220>
<223>  attL recombination site

<400>  13
ctgctttttt atactaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg       60

aacaggtcac tatcagtcaa aatacaatca ttatttgatt tcaattttgt cccactccct      120


<210>  14
<211>  18
<212>  DNA
<213>  Artificial

<220>
<223>  attH4x recombination site

<400>  14
ctttatttca ttaagttg                                                     18


<210>  15
<211>  271
<212>  DNA
<213>  Artificial

<220>
<223>  attP4x recombination site

<400>  15
aattcctctg taacaggtca ctaataccat ctaagtagtt gattcatagt gactgcatat       60

gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga      120

tatttatatc attttacgtt tctcgttcag ctttatttca ttaagttggc attataaaaa      180

agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat acaatcatta      240

tttgatttca attttgtccc actccctccc g                                     271


<210>  16
<211>  121
<212>  DNA
<213>  Artificial

<220>
<223>  attL4x recombination site

<400>  16
ctttatttca ttaagttggc attataaaaa agcattgctt atcaatttgt tgcaacgaac       60

aggtcactat cagtcaaaat acaatcatta tttgatttca attttgtccc actccctccc      120

g                                                                      121


<210>  17
<211>  168
<212>  DNA
<213>  Artificial

<220>
<223>  attR4x recombination site

<400>  17
aattcctctg taacaggtca ctaataccat ctaagtagtt gattcatagt gactgcatat       60

gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga      120

tatttatatc attttacgtt tctcgttcag ctttatttca ttaagttg                   168


<210>  18
<211>  595
<212>  DNA
<213>  Artificial

<220>
<223>  scIHF2

<400>  18
atggctagca ccaagtcaga attgatagaa agacttgcca cccagcaatc gcacattccc       60

gccaagacgg ttgaagatgc agtaaaagag atgctggagc atatggcctc gactcttgcg      120

cagggtggaa gcggcggtct tacaaaagct gaaatgtcag aatatctgtt tgataagctt      180

gggcttagca agcgggatgc caaagaactg gttgaactgt ttttcgaaga gatccgtcgc      240

gctctggaaa acggcgaaca ggtgaaactc tctggttttg gtaacttcga tctgcgtgat      300

aagaatcaac gcccgggacg taacccgaaa acgggcgagg atattcccat tacagcacgg      360

cgcgtggtga ccttcagacc cgggcagaag ttaaaaagcc gggtcgaaaa cgctggtggg      420

ggcgagcgta ttgaaatccg cggtttcggc agtttctctt tgcactaccg cgcaccacgt      480

accggacgta atccgaagac tggcgataaa gtagaactgg aaggaaaata cgttcctcac      540

tttaaacctg gtaaagaact gcgcgatcgc gccaatatct acggtggatc aggct           595


<210>  19
<211>  647
<212>  DNA
<213>  Artificial

<220>
<223>  chloramphenicol resistance gene

<400>  19
atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa       60

cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat      120

attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt      180

cacattcttg cccgcctgat gaatgctcat ccggaattac gtatggcaat gaaagacggt      240

gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa      300

acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat      360

tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag      420

aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg      480

gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc      540

gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat      600

gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggca                    647


<210>  20
<211>  34
<212>  DNA
<213>  Artificial

<220>
<223>  FLP-FRT recombination site

<400>  20
gaagttccta ttctctagaa agtataggaa cttc                                   34


<210>  21
<211>  63
<212>  DNA
<213>  Artificial

<220>
<223>  partial genomic sequence

<400>  21
tcactgccaa aatcgaggcc aattgcaatc gccatcgttt cactccatcc aaaaaaacgg       60

gta                                                                     63


<210>  22
<211>  63
<212>  DNA
<213>  Artificial

<220>
<223>  partial genomic sequence

<400>  22
tacccgtttt tttggatgga gtgaaacgat ggcgattgca attggcctcg attttggcag       60

tga                                                                     63


