LOCUS       sequence-311561-        4031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZFN (left) targeting the AAVS1 locus with obligate heterodimer FokI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4031)
  TITLE     ZFN/TALEN-AAVS1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4031)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pZFN-AAVS1_ELD
FEATURES             Location/Qualifiers
     source          1..4031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     149..169
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     243..262
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        243..261
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             330..395
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             402..422
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1435..1452)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1441..1665
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             1838..2632
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1838..2632
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1892..1911)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2502..2521
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2957..3545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3446..3465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3646..4025
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4026..4031,1..198)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
       61 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      121 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      181 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      241 attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt aagctgatcc
      301 actagtccag tgtggtggaa ttcgccatgg actacaaaga ccatgacggt gattataaag
      361 atcatgacat cgattacaag gatgacgatg acaagatggc ccccaagaag aagaggaagg
      421 tgggcattca cggggtaccc gccgctatgg ccgagcggcc cttccagtgc cggatctgca
      481 tgcggaactt cagctacaac tggcatctgc agcggcacat ccggacccac accggcgaga
      541 agcccttcgc ctgcgacatc tgcggccgga agttcgccag aagcgaccac ctgaccaccc
      601 acaccaagat ccacacaggc agccagaaac cttttcagtg cagaatctgt atgagaaact
      661 tctcccacaa ctacgcccgg gactgccaca tcagaacaca cacaggggag aaaccttttg
      721 cctgcgatat ctgtgggcgc aaatttgccc agaacagcac ccggatcggg cacacaaaga
      781 ttcacctgag aggatcccag ctggtgaaga gcgagctgga ggagaagaag tccgagctgc
      841 ggcacaagct gaagtacgtg ccccacgagt acatcgagct gatcgagatc gccaggaaca
      901 gcacccagga ccgcatcctg gagatgaagg tgatggagtt cttcatgaag gtgtacggct
      961 acaggggaaa gcacctgggc ggaagcagaa agcctgacgg cgccatctat acagtgggca
     1021 gccccatcga ttacggcgtg atcgtggaca caaaggccta cagcggcggc tacaatctgc
     1081 ctatcggcca ggccgacgag atggagagat acgtggagga gaaccagacc cgggataagc
     1141 acctcaaccc caacgagtgg tggaaggtgt accctagcag cgtgaccgag ttcaagttcc
     1201 tgttcgtgag cggccacttc aagggcaact acaaggccca gctgaccagg ctgaaccaca
     1261 tcaccaactg caatggcgcc gtgctgagcg tggaggagct gctgatcggc ggcgagatga
     1321 tcaaagccgg caccctgaca ctggaggagg tgcggcgcaa gttcaacaac ggcgagatca
     1381 acttcagatc ttgataactc gagtctagag ggcccgttta aacccgctga tcagcctcga
     1441 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1501 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1561 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1621 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct actgggcggt
     1681 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
     1741 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     1801 aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     1861 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     1921 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     1981 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     2041 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     2101 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     2161 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     2221 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     2281 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     2341 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     2401 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     2461 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     2521 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     2581 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattattaa
     2641 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2701 catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2761 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatagc
     2821 acgtgctaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3601 acatgttctt gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     3661 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     3721 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     3781 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     3841 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     3901 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     3961 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4021 ccatggtgat g
//
