LOCUS       sequence-307766-        4032 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 4032)
  TITLE     ZFN/TALEN-AAVS1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4032)
  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            "Unpublished"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..4032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        36..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     664..683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        664..682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             751..816
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             823..843
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1856..1873)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1862..2086
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2259..3053
                     /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             2259..3053
                     /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(2313..2332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2923..2942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3379..3967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3868..3887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gactcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      661 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagctgatc
      721 cactagtcca gtgtggtgga attcgccatg gactacaaag accatgacgg tgattataaa
      781 gatcatgaca tcgattacaa ggatgacgat gacaagatgg cccccaagaa gaagaggaag
      841 gtgggcattc acggggtacc cgccgctatg gccgagcggc ccttccagtg ccggatctgc
      901 atgcggaact ttagccagag cagcaacctg gcccggcaca tcagaaccca cacaggcgag
      961 aagcccttcg cctgcgacat ctgcggccgg aagttcgccc ggaccgacta cctggtggac
     1021 cacaccaaga tccacaccgg cagccagaaa ccttttcagt gcagaatctg tatgagaaac
     1081 ttcagctaca acacccacct gaccagacac atccggacac acactgggga gaaacctttt
     1141 gcctgcgata tctgtgggag aaagtttgcc cagggctaca atctggccgg gcacacaaag
     1201 attcacctga gaggatccca gctggtgaag agcgagctgg aggagaagaa gtccgagctg
     1261 cggcacaagc tgaagtacgt gccccacgag tacatcgagc tgatcgagat cgccaggaac
     1321 agcacccagg accgcatcct ggagatgaag gtgatggagt tcttcatgaa ggtgtacggc
     1381 tacaggggaa agcacctggg cggaagcaga aagcctgacg gcgccatcta tacagtgggc
     1441 agccccatcg attacggcgt gatcgtggac acaaaggcct acagcggcgg ctacaatctg
     1501 cctatcggcc aggccgacga gatgcagaga tacgtgaagg agaaccagac ccggaataag
     1561 cacatcaacc ccaacgagtg gtggaaggtg taccctagca gcgtgaccga gttcaagttc
     1621 ctgttcgtga gcggccactt caagggcaac tacaaggccc agctgaccag gctgaaccgc
     1681 aaaaccaact gcaatggcgc cgtgctgagc gtggaggagc tgctgatcgg cggcgagatg
     1741 atcaaagccg gcaccctgac actggaggag gtgcggcgca agttcaacaa cggcgagatc
     1801 aacttcagat cttgataact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     1861 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1921 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     1981 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2041 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tactgggcgg
     2101 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     2161 agccctgcaa agtaaactgg atggctttct cgccgccaag gatctgatgg cgcaggggat
     2221 caagctctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2281 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2341 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2401 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     2461 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2521 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2581 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2641 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2701 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2761 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     2821 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2881 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2941 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3001 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattatta
     3061 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3121 gcatacaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3181 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3241 gcacgtgcta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3301 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3481 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3961 atggaaaaac gccagcaacg cggccttttt acggttcctg ggcttttgct ggccttttgc
     4021 tcacatgttc tt
//
