
                                SEQUENCE LISTING

<110> Seattle Children's Hospital (dba Seattle Children's Research Institute)

<120> SELF-INACTIVATING TRANSPOSASE PLASMIDS
  AND USES THEREOF

<130> SCRI.198WO

<140> PCT/US2019/048663   
<141> 2019-08-28  

<150> 62/724,236          
<151> 2018-08-29  

<160> 20

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 1140
<212> DNA
<213> Artificial Sequence

<220> 
<223> TRANSPOSASE REGION PIGGYBAC TRANSPOSASE 5 PRIME
      CATALYTIC REGION

<400> 1
atgggcagca gcctggacga cgagcacatc ctgtctgccc tgctgcaatc cgacgatgag 60
ctcgtgggcg aggacagcga cagcgagatc agcgatcacg tgtccgagga cgacgtgcag 120
tccgacaccg aggaagcctt catcgacgag gtgcacgaag tgcagcctac cagcagcggc 180
tccgagatcc tggacgagca gaacgtgatc gagcagcccg gaagctccct ggccagcaac 240
agaatcctga ccctgcccca gcggaccatc cggggcaaga acaagcactg ctggtccacc 300
agcaagagca cccggcggtc cagagtgtcc gccctgaata tcgtgcggag ccagaggggc 360
cccacccgga tgtgcagaaa catctacgac cccctgctgt gcttcaagct gttcttcacc 420
gacgagatca tctctgagat cgtgaagtgg accaacgccg agatctccct gaagcggcgc 480
gagtctatga ccggcgccac cttcagagac accaacgagg acgagatcta cgccttcttc 540
ggcatcctcg tgatgaccgc cgtgcggaag gacaaccaca tgagcaccga cgacctgttc 600
gaccggtccc tgagcatggt gtacgtgtcc gtgatgagcc gggacagatt cgacttcctg 660
atccggtgcc tgcggatgga cgacaagagc atcagaccca ccctgcgcga gaacgacgtg 720
ttcacccctg tgcggaagat ctgggacctg ttcatccacc agtgcatcca gaactacacc 780
cctggcgccc acctgaccat cgacgaacag ctgctgggct tcagaggccg gtgccccttc 840
agaatgtaca tccccaacaa gccctctaag tacggcatca agatcctgat gatgtgcgac 900
agcggcacca agtacatgat caacggcatg ccctacctgg gcagaggcac ccagacaaat 960
ggcgtgccac tgggcgagta ctacgtgaaa gaactgagca agcctgtgca cggctcctgc 1020
cggaacatca cctgtgacaa ctggttcacc tccatccccc tggccaagaa tctgctgcag 1080
gaaccctaca agctgacaat cgtgggcacc gtgcggtcca acaagcgcga gattcccgag 1140


<210> 2
<211> 939
<212> DNA
<213> Artifical Sequence

<220> 
<223> EF1a INTRON

<400> 2
gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg 60
ccttgaatta cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt 120
tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg 180
agttgaggcc tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc 240
ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac 300
gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc 360
ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag 420
gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc 480
tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc 540
ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag 600
ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa 660
aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc 720
caggcacctc gattagttcg cttttggagt acgtcgtctt taggttgggg ggaggggttt 780
tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac 840
ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag 900
cctcagacag tggttcaaag tttttttctt ccatttcag                        939

<210> 3
<211> 645
<212> DNA
<213> Artificial Sequence

<220> 
<223> PIGGYBAC TRANSPOSASE 5 PRIME CATALYTIC REGION+STOP
      CODON

<400> 3
gtgctgaaga actcccggtc cagacctgtg ggcaccagca tgttctgctt cgacggccct 60
ctgaccctgg tgtcctacaa gcccaagccc gccaagatgg tgtacctgct gagcagctgt 120
gacgaggacg ccagcatcaa cgagagcacc ggcaagcccc agatggtcat gtactacaac 180
cagaccaagg gcggcgtgga caccctggac cagatgtgca gcgtgatgac atgcagcaga 240
aagaccaacc ggtggcccat ggccctgctg tacggcatga tcaatatcgc ctgcatcaac 300
agcttcatca tctactccca caacgtgtcc agcaagggcg agaaggtgca gagccggaag 360
aaattcatgc ggaacctgta catgagcctg acctccagct tcatgagaaa gcggctggaa 420
gcccccaccc tgaagagata cctgcgggac aacatcagca acatcctgcc caacgaggtg 480
cccggcacca gcgacgatag cacagaggaa cccgtgatga agaagcggac ctactgcacc 540
tactgtccct ctaaaatccg gcggaaggcc aacgccagct gcaaaaagtg caagaaagtg 600
atctgccgcg agcacaacat cgatatgtgc cagagctgct tctga                 645

<210> 4
<211> 134
<212> DNA
<213> Artificial Sequence

<220> 
<223> SV40 LATE POLY(A)

<400> 4
gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata 60
aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg 120
aggtttttta aagc                                                   134

<210> 5
<211> 242
<212> DNA
<213> Artificial Sequence

<220> 
<223> TRANSPOSON REGION 5 PRIME TERMINAL REPEAT
      

<400> 5
ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg cgtaaaattg 60
acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa 120
gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt 180
tatttattta ttaaaaaaaa acaaaaactc aaaatttctt ctataaagta acaaaacttt 240
ta                                                                242

<210> 6
<211> 241
<212> DNA
<213> Artificial Sequence

<220> 
<223> CORE/INSULATOR
      

<400> 6
gagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg 60
gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc 120
ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa 180
cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaggctcgcg 240
a                                                                 241

<210> 7
<211> 245
<212> DNA
<213> Artificial Sequence

<220> 
<223> EF1a PROMOTER

<400> 7
ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg 60
agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa 120
actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt 180
atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac 240
agctg                                                             245

<210> 8
<211> 678
<212> DNA
<213> Artificial Sequence

<220> 
<223> eGFP Flanked by enzymatic cut sites NheI and BamHI

<400> 8
gctagcatgc ccgccatgaa gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag 60
ttcgagctgg tgggcggcgg agagggcacc cccgagcagg gccgcatgac caacaagatg 120
aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac 180
ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc 240
aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac 300
gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc 360
accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc 420
gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc 480
agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc 540
gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag 600
gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc 660
atcgccttcg ccggatcc                                               678

<210> 9
<211> 66
<212> DNA
<213> Artificial Sequence

<220> 
<223> T2A with AGG tag

<400> 9
ggcggcggag agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 60
cctagg                                                            66

<210> 10
<211> 578
<212> DNA
<213> Artificial Sequence

<220> 
<223> DHFRdm+STOP CODON Followed by enzymatic cut sites
      Acc65I and Not I

<400> 10
atggttggtt cgctaaactg catcgtcgct gtgtcccaga acatgggcat cggcaagaac 60
ggggacttcc cctggccacc gctcaggaat gaatccagat atttccagag aatgaccaca 120
acctcttcag tagaaggtaa acagaatctg gtgattatgg gtaagaagac ctggttctcc 180
attcctgaga agaatcgacc tttaaagggt agaattaatt tagttctcag cagagaactc 240
aaggaacctc cacaaggagc tcattttctt tccagaagtc tagatgatgc cttaaaactt 300
actgaacaac cagaattagc aaataaagta gacatggtct ggatagttgg tggcagttct 360
gtttataagg aagccatgaa tcacccaggc catcttaaac tatttgtgac aaggatcatg 420
caagactttg aaagtgacac gttttttcca gaaattgatt tggagaaata taaacttctg 480
ccagaatacc caggtgttct ctctgatgtc caggaggaga aaggcattaa gtacaaattt 540
gaagtatatg agaagaatga ttaaggtacc gcggccgc                         578

<210> 11
<211> 135
<212> DNA
<213> Artificial Sequence

<220> 
<223> RBG/POLY(A)

<400> 11
ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg 60
ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact 120
cggaaggaca taagg                                                  135

<210> 12
<211> 243
<212> DNA
<213> Artificial Sequence

<220> 
<223> CORE/INSULATOR

<400> 12
atggctagat ctttttcccc gtatcccccc aggtgtctgc aggctcaaag agcagcgaga 60
agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt ccccgcacgc 120
tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg cggctcgctg 180
ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg gggggctgtc 240
cct                                                               243

<210> 13
<211> 313
<212> DNA
<213> Artificial Sequence

<220> 
<223> 3 PRIME TERMINAL REPEAT

<400> 13
gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata 60
acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca 120
ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac 180
gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg 240
ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc 300
tttctagggt taa                                                    313

<210> 14
<211> 43
<212> DNA
<213> Artificial Sequence

<220> 
<223> 3 PRIME TERMINAL REPEATNANOPLASMID BACKBONE
      TRPA TERMINATOR Preceded by enzymatic cut site
      SalI

<400> 14
gtcgacccgc ctaatgagcc gcctaatgag cgggcttttt ttt                   43

<210> 15
<211> 281
<212> DNA
<213> Artificial Sequence

<220> 
<223> R6K MINI ORIGIN

<400> 15
ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt 60
ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt 120
gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc 180
ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat 240
gagagcttag tacgtactat caacaggttg aactgctgat c                     281

<210> 16
<211> 150
<212> DNA
<213> Artificial Sequence

<220> 
<223> RNA-OUT

<400> 16
cacgttgtgg tagaattggt aaagagagtc gtgtaaaata tcgagttcgc acatcttgtt 60
gtctgattat tgatttttgg cgaaaccatt tgatcatatg acaagatgtg tatctacctt 120
aacttaatga ttttgataaa aatcattagg                                  150

<210> 17
<211> 467
<212> DNA
<213> Artificial Sequence

<220> 
<223> CMV-ENHANCER

<400> 17
ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 60
gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 120
tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 180
aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 240
caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 300
acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 360
ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 420
gatttccaag tctccacccc attgacgtca atgggagttt gttttgg               467

<210> 18
<211> 163
<212> DNA
<213> Artificial Sequence

<220> 
<223> CMV-CORE PROMOTER

<400> 18
caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 60
ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag 120
atcgcctgga gacgccatcc acgctgtttt gacctccata gaa                   163

<210> 19
<211> 993
<212> DNA
<213> Artificial Sequence

<220> 
<223> SUICIDE GENE SR39TK
      

<400> 19
atgcccacgc tactgcgggt ttatatagac ggtccccacg ggatggggaa aaccaccacc 60
acgcaactgc tggtggccct gggttcgcgc gacgatatcg tctacgtacc cgagccgatg 120
acttactggc gggtgctggg ggcttccgag acaatcgcga acatctacac cacacaacac 180
cgcctcgacc agggtgagat atcggccggg gacgcggcgg tggtaatgac aagcgcccag 240
ataacaatgg gcatgcctta tgccgtgacc gacgccgttc tggctcctca tatcgggggg 300
gaggctggga gctcacatgc cccgcccccg gccctcacca tcttcctcga ccgccatccc 360
atcgccttca tgctgtgcta cccggccgcg cggtacctta tgggcagcat gaccccccag 420
gccgtgctgg cgttcgtggc cctcatcccg ccgaccttgc ccggcaccaa catcgtgctt 480
ggggcccttc cggaggacag acacatcgac cgcctggcca aacgccagcg ccccggcgag 540
cggctggacc tggctatgct ggctgcgatt cgccgcgttt acgggctact tgccaatacg 600
gtgcggtatc tgcagtgcgg cgggtcgtgg cgggaggact ggggacagct ttcggggacg 660
gccgtgccgc cccagggtgc cgagccccag agcaacgcgg gcccacgacc ccatatcggg 720
gacacgttat ttaccctgtt tcgggccccc gagttgctgg cccccaacgg cgacctgtat 780
aacgtgtttg cctgggcctt ggacgtcttg gccaaacgcc tccgttccat gcacgtcttt 840
atcctggatt acgaccaatc gcccgccggc tgccgggacg ccctgctgca acttacctcc 900
gggatggtcc agacccacgt caccaccccc ggctccatac cgacgatatg cgacctggcg 960
cgcacgtttg cccgggagat gggggaggct aac                              993

<210> 20
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> T2A

<400> 20
ggcggcggag agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 60
cct                                                               63
