LOCUS       sequence-311594-        4031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZFN (right) 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  JOURNAL   Unpublished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            "Unpublished"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-KKR
FEATURES             Location/Qualifiers
     source          1..4031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        140..343
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     294..314
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     388..407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             475..540
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             547..567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1580..1597)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1586..1810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             1983..2777
                     /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             1983..2777
                     /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(2037..2056)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2647..2666
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3102..3690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3591..3610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(3791..4031,1..139)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
       61 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      121 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      181 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      241 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      301 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      361 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      421 ctagcgttta aacttaagct gatccactag tccagtgtgg tggaattcgc catggactac
      481 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
      541 atggccccca agaagaagag gaaggtgggc attcacgggg tacccgccgc tatggccgag
      601 cggcccttcc agtgccggat ctgcatgcgg aactttagcc agagcagcaa cctggcccgg
      661 cacatcagaa cccacacagg cgagaagccc ttcgcctgcg acatctgcgg ccggaagttc
      721 gcccggaccg actacctggt ggaccacacc aagatccaca ccggcagcca gaaacctttt
      781 cagtgcagaa tctgtatgag aaacttcagc tacaacaccc acctgaccag acacatccgg
      841 acacacactg gggagaaacc ttttgcctgc gatatctgtg ggagaaagtt tgcccagggc
      901 tacaatctgg ccgggcacac aaagattcac ctgagaggat cccagctggt gaagagcgag
      961 ctggaggaga agaagtccga gctgcggcac aagctgaagt acgtgcccca cgagtacatc
     1021 gagctgatcg agatcgccag gaacagcacc caggaccgca tcctggagat gaaggtgatg
     1081 gagttcttca tgaaggtgta cggctacagg ggaaagcacc tgggcggaag cagaaagcct
     1141 gacggcgcca tctatacagt gggcagcccc atcgattacg gcgtgatcgt ggacacaaag
     1201 gcctacagcg gcggctacaa tctgcctatc ggccaggccg acgagatgca gagatacgtg
     1261 aaggagaacc agacccggaa taagcacatc aaccccaacg agtggtggaa ggtgtaccct
     1321 agcagcgtga ccgagttcaa gttcctgttc gtgagcggcc acttcaaggg caactacaag
     1381 gcccagctga ccaggctgaa ccgcaaaacc aactgcaatg gcgccgtgct gagcgtggag
     1441 gagctgctga tcggcggcga gatgatcaaa gccggcaccc tgacactgga ggaggtgcgg
     1501 cgcaagttca acaacggcga gatcaacttc agatcttgat aactcgagtc tagagggccc
     1561 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1621 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1681 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1741 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1801 ggctctatgg cttctactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     1861 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     1921 caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc
     1981 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2041 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2101 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2161 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2221 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2281 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2341 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2401 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2461 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     2521 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2581 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2641 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2701 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2761 ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta
     2821 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     2881 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2941 ccctgataaa tgcttcaata atagcacgtg ctaaaacttc atttttaatt taaaaggatc
     3001 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3061 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3181 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3241 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3301 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3361 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3421 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3481 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3541 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3601 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3661 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3721 ctggcctttt gctggccttt tgctcacatg ttcttgctgc ttcgcgatgt acgggccaga
     3781 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     3841 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     3901 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     3961 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4021 gcagtacatc a
//
