LOCUS       sequence-311595-        7517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN (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 7517)
  TITLE     ZFN/TALEN-AAVS1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7517)
  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            "Unpublished"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: pCMV-AAVS1-TALEN-L
FEATURES             Location/Qualifiers
     source          1..7517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             669..734
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             741..761
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3705..3723)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3706..3723)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3743..3760)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3749..3973
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4019..4447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4106..4125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4316..4337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4456..4476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4461..4790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4641..4776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4703..4722
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4956..5077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4993..5012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5047..5066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5126..5142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5126..5142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5139..5161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5150..5166
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5174..5180,5181..5198,5199..5204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5219..5240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5357..5374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5528..6116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5608..5627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6287..7078,7079..7147))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6910..6929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7148..7252)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7327..7346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ccaccatgga ctacaaggac cacgacggcg attacaagga tcacgatatc gactataagg
      721 acgacgatga caagatggcc cccaagaaaa agcggaaagt gggcatccac cggggggtac
      781 caatggtgga cctgcggacc ctgggctaca gccagcagca gcaagagaag atcaagccca
      841 aagtgcggag caccgtggcc cagcaccacg aagccctcgt gggccacggc tttacccacg
      901 cccacatcgt ggccctgagc cagcatcctg ccgccctggg aacagtggcc gtgaagtacc
      961 aggacatgat cgccgccctg cccgaggcca cacacgaggc cattgtggga gtgggcaagc
     1021 agtggtccgg cgccagagcc ctggaagccc tgctgacagt ggccggcgag ctgagaggac
     1081 cccccctgca gctggatacc ggccagctgc tgaagatcgc caagcgggga ggcgtgaccg
     1141 ccgtggaagc tgtgcacgct tggcggaacg ccctgacagg cgcccctctg aacctgaccc
     1201 ccgaccaggt ggtggccatt gcctctcacg acggcggcaa gcaggccctg gaaaccgtgc
     1261 agagactgct gcccgtgctg tgccaggacc acggcctgac acctgagcag gtcgtggcta
     1321 tcgccagcca cgatggggga aaacaggctc tggaaacagt ccagcggctg ctccctgtcc
     1381 tgtgtcaggc ccatggactg accccagatc aggtcgtcgc aatcgcctcc catgacgggg
     1441 ggaaacaggc ccttgagaca gtgcaacgcc tcctgccagt gctgtgtcaa gctcacgggc
     1501 tgacccctgc acaggtggtc gctattgcta gccatgacgg cggaaagcaa gcactcgaaa
     1561 ctgttcagcg cctgttgcca gtcttgtgtc aggatcatgg cctcacacca gaccaagtgg
     1621 tggcaatcgc tagcaacggc ggaggaaagc aagctcttga gactgtgcag aggctgttgc
     1681 cggttctctg tcaagaccac ggactgactc ccgaacaagt ggtcgccata gcttcccatg
     1741 atgggggcaa gcaggcattg gagactgtgc aacggctgct cccggtgctt tgccaggctc
     1801 acggactcac tccagaccag gttgtggcta tagcaagtca tgatggcggc aaacaagctc
     1861 ttgaaaccgt tcaacggctt ctgcctgtgc tctgccaagc acatggcctg actcccgccc
     1921 aggttgtcgc aattgccagc aacatcggag ggaaacaagc cttggaaact gtccaaagac
     1981 tgcttccggt cctttgtcaa gatcatggac ttacgccgga tcaagtggtt gctatcgcat
     2041 cccacgatgg cgggaagcag gcacttgaaa cggtccaaag gctcctgccg gttttgtgcc
     2101 aagaccatgg gctcacaccc gaacaggttg tcgctatcgc ttcacacgac ggtggcaaac
     2161 aggcattgga aacagttcag agactccttc ctgttctttg ccaagcccac ggccttaccc
     2221 ccgatcaagt tgtggcaatt gcaagccacg acggggggaa gcaagctctg gaaacggttc
     2281 aaagattgct gccagtcctg tgccaggcac atggccttac ccctgcacaa gtcgtggcta
     2341 tagcctcaca tgacggtgga aaacaagccc tggaaacggt gcaacgactt ctcccagttc
     2401 tgtgccagga tcatgggctg actccggatc aggttgtggc cattgcttcc aatatcggag
     2461 ggaagcaggc tctcgaaacc gtgcaacgcc tgcttcccgt cttgtgccaa gatcacggtc
     2521 ttactccaga gcaagtcgtt gccattgcat cccacgacgg gggcaaacaa gccctcgaaa
     2581 ctgtccagcg gctcctgccc gttctttgtc aagcacacgg attgacacct gaccaagttg
     2641 ttgctattgc ctccaacatt ggtggcaagc aagccttgga gacagttcaa aggctcttgc
     2701 ccgtgctctg tcaggctcat ggtcttacac ccgctcaggt ggtggcaata gccaacaaca
     2761 acggcggaaa acaggcactt gagactgtcc agagattgct cccagtgttg tgtcaagacc
     2821 acggcttgac accagaacag gtggtcgcaa tagcatccaa cggcggaggc agacccgccc
     2881 tggaatccat tgtggcccag ctgagcagac ccgacccagc cctggccgcc ctgaccaacg
     2941 atcatctggt ggccctggcc tgcctgggcg gcagacctgc tctggacgcc gtgaagaagg
     3001 gcctgcctca cgcccctgcc ctgatcaagc ggaccaacag aagaatcccc gagcggacca
     3061 gccaccgggt ggccggatcc cagctggtca agagcgagct ggaagagaag aagtccgagc
     3121 tgcggcacaa gctgaaatac gtgccccacg agtacatcga gctgatcgag atcgcccgga
     3181 acagcaccca ggacagaatc ctggaaatga aggtcatgga attcttcatg aaggtgtacg
     3241 gctaccgggg caagcacctg ggcggctcca gaaagcccga cggcgccatc tacaccgtgg
     3301 gcagccccat cgactacggc gtgatcgtgg acaccaaggc ctacagcggc ggctacaacc
     3361 tgcccatcgg acaggccgac gagatggaaa gatacgtgga agagaaccag accagggaca
     3421 agcacctgaa ccccaacgag tggtggaagg tgtaccccag cagcgtgacc gagttcaagt
     3481 tcctgttcgt gtccggccac ttcaagggca actacaaagc ccagctgacc cggctgaacc
     3541 acatcaccaa ctgcaacggc gccgtgctga gcgtggaaga actgctgatc ggcggcgaga
     3601 tgatcaaggc cggcaccctg accctggaag aagtgcggcg gaagttcaac aatggcgaga
     3661 tcaacttccg cagctgactc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     3721 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3781 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3841 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3901 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3961 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4021 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4081 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4141 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4201 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4261 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4321 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4381 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4441 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4501 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4621 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4681 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4741 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
     4801 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4861 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4921 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4981 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5041 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5101 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5161 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5221 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5281 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5341 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7261 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     7321 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     7381 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     7441 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     7501 ggccagatat acgcgtt
//
