LOCUS       sequence-307767-        7518 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 7518)
  TITLE     ZFN/TALEN-AAVS1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7518)
  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            "Unpublished"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..7518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             903..968
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             975..995
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3939..3957)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3940..3957)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3977..3994)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3983..4207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4253..4681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4340..4359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4550..4571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4690..4710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4695..5025
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4876..5011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4938..4957
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5191..5312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5228..5247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5282..5301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5361..5377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5361..5377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5374..5396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5385..5401
                     /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(5409..5415,5416..5433,5434..5439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5454..5475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5592..5609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5763..6351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5843..5862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6522..7313,7314..7382))
                     /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     7145..7164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7383..7487)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttccacca
      901 tggactacaa ggaccacgac ggcgattaca aggatcacga tatcgactat aaggacgacg
      961 atgacaagat ggcccccaag aaaaagcgga aagtgggcat ccaccggggg gtaccaatgg
     1021 tggacctgcg gaccctgggc tacagccagc agcagcaaga gaagatcaag cccaaagtgc
     1081 ggagcaccgt ggcccagcac cacgaagccc tcgtgggcca cggctttacc cacgcccaca
     1141 tcgtggccct gagccagcat cctgccgccc tgggaacagt ggccgtgaag taccaggaca
     1201 tgatcgccgc cctgcccgag gccacacacg aggccattgt gggagtgggc aagcagtggt
     1261 ccggcgccag agccctggaa gccctgctga cagtggccgg cgagctgaga ggaccccccc
     1321 tgcagctgga taccggccag ctgctgaaga tcgccaagcg gggaggcgtg accgccgtgg
     1381 aagctgtgca cgcttggcgg aacgccctga caggcgcccc tctgaacctg acccccgacc
     1441 aggtggtggc cattgcctct cacgacggcg gcaagcaggc cctggaaacc gtgcagagac
     1501 tgctgcccgt gctgtgccag gaccacggcc tgacacctga gcaggtcgtg gctatcgcca
     1561 gccacgatgg gggaaaacag gctctggaaa cagtccagcg gctgctccct gtcctgtgtc
     1621 aggcccatgg actgacccca gatcaggtcg tcgcaatcgc ctcccatgac ggggggaaac
     1681 aggcccttga gacagtgcaa cgcctcctgc cagtgctgtg tcaagctcac gggctgaccc
     1741 ctgcacaggt ggtcgctatt gctagccatg acggcggaaa gcaagcactc gaaactgttc
     1801 agcgcctgtt gccagtcttg tgtcaggatc atggcctcac accagaccaa gtggtggcaa
     1861 tcgctagcaa cggcggagga aagcaagctc ttgagactgt gcagaggctg ttgccggttc
     1921 tctgtcaaga ccacggactg actcccgaac aagtggtcgc catagcttcc catgatgggg
     1981 gcaagcaggc attggagact gtgcaacggc tgctcccggt gctttgccag gctcacggac
     2041 tcactccaga ccaggttgtg gctatagcaa gtcatgatgg cggcaaacaa gctcttgaaa
     2101 ccgttcaacg gcttctgcct gtgctctgcc aagcacatgg cctgactccc gcccaggttg
     2161 tcgcaattgc cagcaacatc ggagggaaac aagccttgga aactgtccaa agactgcttc
     2221 cggtcctttg tcaagatcat ggacttacgc cggatcaagt ggttgctatc gcatcccacg
     2281 atggcgggaa gcaggcactt gaaacggtcc aaaggctcct gccggttttg tgccaagacc
     2341 atgggctcac acccgaacag gttgtcgcta tcgcttcaca cgacggtggc aaacaggcat
     2401 tggaaacagt tcagagactc cttcctgttc tttgccaagc ccacggcctt acccccgatc
     2461 aagttgtggc aattgcaagc cacgacgggg ggaagcaagc tctggaaacg gttcaaagat
     2521 tgctgccagt cctgtgccag gcacatggcc ttacccctgc acaagtcgtg gctatagcct
     2581 cacatgacgg tggaaaacaa gccctggaaa cggtgcaacg acttctccca gttctgtgcc
     2641 aggatcatgg gctgactccg gatcaggttg tggccattgc ttccaatatc ggagggaagc
     2701 aggctctcga aaccgtgcaa cgcctgcttc ccgtcttgtg ccaagatcac ggtcttactc
     2761 cagagcaagt cgttgccatt gcatcccacg acgggggcaa acaagccctc gaaactgtcc
     2821 agcggctcct gcccgttctt tgtcaagcac acggattgac acctgaccaa gttgttgcta
     2881 ttgcctccaa cattggtggc aagcaagcct tggagacagt tcaaaggctc ttgcccgtgc
     2941 tctgtcaggc tcatggtctt acacccgctc aggtggtggc aatagccaac aacaacggcg
     3001 gaaaacaggc acttgagact gtccagagat tgctcccagt gttgtgtcaa gaccacggct
     3061 tgacaccaga acaggtggtc gcaatagcat ccaacggcgg aggcagaccc gccctggaat
     3121 ccattgtggc ccagctgagc agacccgacc cagccctggc cgccctgacc aacgatcatc
     3181 tggtggccct ggcctgcctg ggcggcagac ctgctctgga cgccgtgaag aagggcctgc
     3241 ctcacgcccc tgccctgatc aagcggacca acagaagaat ccccgagcgg accagccacc
     3301 gggtggccgg atcccagctg gtcaagagcg agctggaaga gaagaagtcc gagctgcggc
     3361 acaagctgaa atacgtgccc cacgagtaca tcgagctgat cgagatcgcc cggaacagca
     3421 cccaggacag aatcctggaa atgaaggtca tggaattctt catgaaggtg tacggctacc
     3481 ggggcaagca cctgggcggc tccagaaagc ccgacggcgc catctacacc gtgggcagcc
     3541 ccatcgacta cggcgtgatc gtggacacca aggcctacag cggcggctac aacctgccca
     3601 tcggacaggc cgacgagatg gaaagatacg tggaagagaa ccagaccagg gacaagcacc
     3661 tgaaccccaa cgagtggtgg aaggtgtacc ccagcagcgt gaccgagttc aagttcctgt
     3721 tcgtgtccgg ccacttcaag ggcaactaca aagcccagct gacccggctg aaccacatca
     3781 ccaactgcaa cggcgccgtg ctgagcgtgg aagaactgct gatcggcggc gagatgatca
     3841 aggccggcac cctgaccctg gaagaagtgc ggcggaagtt caacaatggc gagatcaact
     3901 tccgcagctg actcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     3961 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4021 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4081 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4141 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4201 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4261 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4321 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4381 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     4441 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4501 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4561 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4621 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4681 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc
     4741 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4801 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4861 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4921 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4981 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccgaaatg
     5041 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5101 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5161 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5341 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5401 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5461 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5521 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5941 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6181 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7501 aaagtgccac ctgacgtc
//
