LOCUS       sequence-307768-        7520 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 7520)
  TITLE     ZFN/TALEN-AAVS1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7520)
  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            "Unpublished"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..7520
                     /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(3941..3959)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3942..3959)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3979..3996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3985..4209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4255..4683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4342..4361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4552..4573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4692..4712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4697..5027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4878..5013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4940..4959
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5193..5314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5230..5249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5284..5303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5363..5379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5363..5379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5376..5398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5387..5403
                     /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(5411..5417,5418..5435,5436..5441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5456..5477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5594..5611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5765..6353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5845..5864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6524..7315,7316..7384))
                     /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     7147..7166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7385..7489)
                     /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 cattgccagc aacggcggag gcaagcaggc cctggaaacc gtgcagagac
     1501 tgctgcccgt gctgtgccag gaccacggcc tgacacctga gcaggtcgtg gctatcgcct
     1561 ccaatggcgg cggaaaacag gctctggaaa cagtccagcg gctgctccct gtcctgtgtc
     1621 aggcccatgg actgacccca gatcaggtcg tcgcaatcgc tagcaatggt ggcgggaagc
     1681 aggcacttga gacagtgcaa cggctcctgc cagtgctgtg tcaagctcac gggctgaccc
     1741 ctgcacaggt ggtcgctatc gcaagccacg atggcggcaa acaagccctt gagactgtcc
     1801 agcgcctgtt gccagtcttg tgccaggatc atgggctgac tcccgatcaa gtggttgcta
     1861 ttgcctctaa tgggggagga aagcaagcac tcgaaactgt gcaacgcctg ctgcctgtcc
     1921 tctgccaaga tcacggactc acacccgatc aggttgtggc aatagccaac aacaatgggg
     1981 ggaaacaggc cttggagact gttcagaggc tcttgcccgt gctctgtcag gcacatggcc
     2041 tcacgcctga ccaagtggtg gcaatcgctt ccaacggggg tggcaagcaa gctcttgaaa
     2101 cagttcaaag gctgttgccg gttctgtgcc aagcacacgg cctcactcca gcccaggttg
     2161 tcgccattgc ttctcacgac gggggaaagc aggctctcga aaccgttcaa cgattgcttc
     2221 ccgttctctg tcaagaccat gggttgacac cagaccaagt cgtggccata gcaagcaata
     2281 tcgggggcaa acaagcactg gaaacggtcc aaagactcct cccggtgctt tgtcaagatc
     2341 atggccttac ccccgagcaa gttgtcgcaa ttgccagcca tgatggtgga aaacaggccc
     2401 tcgaaacggt gcaaaggctt ctcccagttc tctgccaagc tcatggtctt acgccggatc
     2461 aagtcgtcgc tatagcatcc catgacgggg ggaagcaagc cttggaaacg gtgcagcggt
     2521 tgctgccggt gttgtgtcag gcacacggat tgacaccagc acaggttgtg gctattgcat
     2581 ccaacatcgg cgggaagcaa gctctggaaa ccgtgcaacg gcttttgcca gtcctttgcc
     2641 aagaccatgg acttactccg gaccaggtcg tggcaattgc ttctaatatt ggcggaaagc
     2701 aggcccttga aactgttcag cggctgcttc cggtgctttg ccaggatcac gggttgaccc
     2761 cggaacaggt tgtggccata gcttccaatg gcggagggaa acaagccctc gaaacagtcc
     2821 agagattgct cccagtgttg tgccaagccc atgggctcac acctgatcaa gttgtcgcta
     2881 tcgcctcaca cgacggtggc aaacaggcat tggagacagt ccaacgtctg cttccagttc
     2941 tgtgccaggc tcacggactt accccagccc aagttgttgc aatcgcatcc cacgatgggg
     3001 gcaagcaggc attggaaact gtgcaacgat tgctccctgt gctttgtcag gaccacgggc
     3061 tcacacccga acaagtcgtt gcaattgcta gtaatggcgg 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 cagagatacg tgaaagagaa ccagacccgg aacaagcaca
     3661 tcaaccccaa cgagtggtgg aaggtgtacc ccagcagcgt gaccgagttc aagttcctgt
     3721 tcgtgtccgg ccacttcaag ggcaactaca aagcccagct gacccggctg aacagaaaga
     3781 ccaactgcaa cggcgccgtg ctgagcgtgg aagaactgct gatcggcggc gagatgatca
     3841 aggccggcac cctgaccctg gaagaagtgc ggcggaagtt caacaacggc gagatcaact
     3901 tctgagcggc cgcctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     3961 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4021 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4081 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4141 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4201 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     4261 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4321 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4381 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4441 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4501 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4561 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4621 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4681 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag
     4741 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4801 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4861 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4921 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4981 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctccccgaaa
     5041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5941 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6181 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc acctgacgtc
//
