LOCUS       sequence-447560-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 TALEN expression vector (Left).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Oceguera-Yanez F, Kim SI, Matsumoto T, Tan GW, Xiang L, Hatani T, 
            Kondo T, Ikeya M, Yoshida Y, Inoue H, Woltjen K
  TITLE     Engineering the AAVS1 locus for consistent and scalable transgene 
            expression in human iPSCs and their differentiated derivatives.
  JOURNAL   Methods. 2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 
            10.1016/j.ymeth.2015.12.012.
  PUBMED    26707206
REFERENCE   2  (bases 1 to 5863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods. 
            2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 
            10.1016/j.ymeth.2015.12.012."
COMMENT     Sequence Label: pTALdNC-AAVS1_T1
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             505..525
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             526..552
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             3475..4062
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     protein_bind    complement(4092..4191)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4303..4322)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4314..5123
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4381..4400)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5213..5801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5702..5721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatcacca tggcttcctc ccctccaaag aaaaagagaa aggttagttg gaaggacgca
      541 agtggttggt ctagaatgca tgcggccccg cgacggcgtg ctgcgcaacc ctccgacgct
      601 tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca gcaagagaag
      661 atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt gggccatggg
      721 tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg gaccgtcgct
      781 gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga catcgttggc
      841 gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga tgcgggggag
      901 ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc aaaacgtggc
      961 ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg tgcccccctg
     1021 aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc
     1081 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa
     1141 gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt gcagcggctg
     1201 ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc
     1261 cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag
     1321 gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa
     1381 gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg
     1441 gaccaagtgg tggctatcgc cagcaacaat ggcggcaagc aagcgctcga aacggtgcag
     1501 cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc
     1561 gccagcaacg gtggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg
     1621 tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca cgatggcggc
     1681 aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg
     1741 actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg
     1801 gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgactccgga ccaagtggtg
     1861 gctatcgcca gccacgatgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg
     1921 gtgctgtgcc aggaccatgg cctgactccg gaccaagtgg tggctatcgc cagccacgat
     1981 ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat
     2041 ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa gcaagcgctc
     2101 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac tccggaccaa
     2161 gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg
     2221 ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc
     2281 cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag
     2341 gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacattgg cggcaagcaa
     2401 gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgactccg
     2461 gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga aacggtgcag
     2521 cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt ggtggctatc
     2581 gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg
     2641 tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca cgatggcggc
     2701 aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg
     2761 actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg
     2821 gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg
     2881 gctatcgcca gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg
     2941 gtgctgtgcc aggaccatgg cctgactccg gaccaagtgg tggctatcgc cagccacgat
     3001 ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat
     3061 ggcctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc
     3121 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa
     3181 gtggtggcta tcgccagcaa caatggcggc aagcaagcgc tcgaaacggt gcagcggctg
     3241 ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc
     3301 aacggtggcg gcaagcaagc gctcgaaagc attgtggccc agctgagccg gcctgatccg
     3361 gcgttggccg cgttgaccaa cgaccacctc gtcgccttgg cctgcctcgg cggacgtcct
     3421 gccatggatg cagtgaaaaa gggattgccg cacgcgccgg aattgatcag atcccagcta
     3481 gtgaaatctg aattggaaga gaagaaatct gaacttagac ataaattgaa atatgtgcca
     3541 catgaatata ttgaattgat tgaaatcgca agaaattcaa ctcaggatag aatccttgaa
     3601 atgaaggtga tggagttctt tatgaaggtt tatggttatc gtggtaaaca tttgggtgga
     3661 tcaaggaaac cagacggagc aatttatact gtcggatctc ctattgatta cggtgtgatc
     3721 gttgatacta aggcatattc aggaggttat aatcttccaa ttggtcaagc agatgaaatg
     3781 caaagatatg tcgaagagaa tcaaacaaga aacaagcata tcaaccctaa tgaatggtgg
     3841 aaagtctatc catcttcagt aacagaattt aagttcttgt ttgtgagtgg tcatttcaaa
     3901 ggaaactaca aagctcagct tacaagattg aatcatatca ctaattgtaa tggagctgtt
     3961 cttagtgtag aagagctttt gattggtgga gaaatgatta aagctggtac attgacactt
     4021 gaggaagtga gaaggaaatt taataacggt gagataaact tttaataggc cgccgcactc
     4081 gagatatcta gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     4141 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4201 tgcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4261 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     4321 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     4381 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     4441 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     4501 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     4561 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     4621 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     4681 cagtgtccct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     4741 gcgtatttcg tctggctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     4801 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     4861 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     4921 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     4981 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     5041 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     5101 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacat tattcagatt
     5161 gggccccgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5821 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
