LOCUS       sequence-311596-        7519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN (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 7519)
  TITLE     ZFN/TALEN-AAVS1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7519)
  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            "Unpublished"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-R
FEATURES             Location/Qualifiers
     source          1..7519
                     /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(3707..3725)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3708..3725)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3745..3762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3751..3975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4021..4449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4108..4127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4318..4339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4458..4478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4463..4792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4643..4778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4705..4724
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4958..5079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4995..5014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5049..5068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5128..5144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5128..5144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5141..5163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5152..5168
                     /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(5176..5182,5183..5200,5201..5206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5221..5242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5359..5376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5530..6118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5610..5629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6289..7080,7081..7149))
                     /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     6912..6931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7150..7254)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7329..7348)
                     /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 gccagcaacg gcggaggcaa gcaggccctg gaaaccgtgc
     1261 agagactgct gcccgtgctg tgccaggacc acggcctgac acctgagcag gtcgtggcta
     1321 tcgcctccaa tggcggcgga aaacaggctc tggaaacagt ccagcggctg ctccctgtcc
     1381 tgtgtcaggc ccatggactg accccagatc aggtcgtcgc aatcgctagc aatggtggcg
     1441 ggaagcaggc acttgagaca gtgcaacggc tcctgccagt gctgtgtcaa gctcacgggc
     1501 tgacccctgc acaggtggtc gctatcgcaa gccacgatgg cggcaaacaa gcccttgaga
     1561 ctgtccagcg cctgttgcca gtcttgtgcc aggatcatgg gctgactccc gatcaagtgg
     1621 ttgctattgc ctctaatggg ggaggaaagc aagcactcga aactgtgcaa cgcctgctgc
     1681 ctgtcctctg ccaagatcac ggactcacac ccgatcaggt tgtggcaata gccaacaaca
     1741 atggggggaa acaggccttg gagactgttc agaggctctt gcccgtgctc tgtcaggcac
     1801 atggcctcac gcctgaccaa gtggtggcaa tcgcttccaa cgggggtggc aagcaagctc
     1861 ttgaaacagt tcaaaggctg ttgccggttc tgtgccaagc acacggcctc actccagccc
     1921 aggttgtcgc cattgcttct cacgacgggg gaaagcaggc tctcgaaacc gttcaacgat
     1981 tgcttcccgt tctctgtcaa gaccatgggt tgacaccaga ccaagtcgtg gccatagcaa
     2041 gcaatatcgg gggcaaacaa gcactggaaa cggtccaaag actcctcccg gtgctttgtc
     2101 aagatcatgg ccttaccccc gagcaagttg tcgcaattgc cagccatgat ggtggaaaac
     2161 aggccctcga aacggtgcaa aggcttctcc cagttctctg ccaagctcat ggtcttacgc
     2221 cggatcaagt cgtcgctata gcatcccatg acggggggaa gcaagccttg gaaacggtgc
     2281 agcggttgct gccggtgttg tgtcaggcac acggattgac accagcacag gttgtggcta
     2341 ttgcatccaa catcggcggg aagcaagctc tggaaaccgt gcaacggctt ttgccagtcc
     2401 tttgccaaga ccatggactt actccggacc aggtcgtggc aattgcttct aatattggcg
     2461 gaaagcaggc ccttgaaact gttcagcggc tgcttccggt gctttgccag gatcacgggt
     2521 tgaccccgga acaggttgtg gccatagctt ccaatggcgg agggaaacaa gccctcgaaa
     2581 cagtccagag attgctccca gtgttgtgcc aagcccatgg gctcacacct gatcaagttg
     2641 tcgctatcgc ctcacacgac ggtggcaaac aggcattgga gacagtccaa cgtctgcttc
     2701 cagttctgtg ccaggctcac ggacttaccc cagcccaagt tgttgcaatc gcatcccacg
     2761 atgggggcaa gcaggcattg gaaactgtgc aacgattgct ccctgtgctt tgtcaggacc
     2821 acgggctcac acccgaacaa gtcgttgcaa ttgctagtaa tggcggaggc 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 gagatgcaga gatacgtgaa agagaaccag acccggaaca
     3421 agcacatcaa ccccaacgag tggtggaagg tgtaccccag cagcgtgacc gagttcaagt
     3481 tcctgttcgt gtccggccac ttcaagggca actacaaagc ccagctgacc cggctgaaca
     3541 gaaagaccaa ctgcaacggc gccgtgctga gcgtggaaga actgctgatc ggcggcgaga
     3601 tgatcaaggc cggcaccctg accctggaag aagtgcggcg gaagttcaac aacggcgaga
     3661 tcaacttctg agcggccgcc tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     3721 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3781 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3841 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3901 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3961 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     4021 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4081 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4141 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4201 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4261 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4321 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4381 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4441 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4561 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4621 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4681 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4741 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4801 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4861 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4921 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5101 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5161 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5221 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5281 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5341 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5401 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5461 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5521 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5581 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5641 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5701 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5761 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5821 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5881 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5941 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6001 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6061 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6121 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6181 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6241 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6301 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6361 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6421 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6481 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6541 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6601 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6661 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6721 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6781 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6841 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6901 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6961 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7021 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7081 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7141 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7201 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7261 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     7321 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7381 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7441 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     7501 cgggccagat atacgcgtt
//
