LOCUS       sequence-425811-        6197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for knocking in an N-terminal tag (FKBP12F36V and V5) for
            human TWIST1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  AUTHORS   Kim S, Morgunova E, Naqvi S, Goovaerts S, Bader M, Koska M, Popov A,
            Luong C, Pogson A, Swigut T, Claes P, Taipale J, Wysocka J
  TITLE     DNA-guided transcription factor cooperativity shapes face and limb 
            mesenchyme.
  JOURNAL   Cell. 2024 Feb 1;187(3):692-711.e26. doi: 
            10.1016/j.cell.2023.12.032. Epub 2024 Jan 22.
  PUBMED    38262408
REFERENCE   2  (bases 1 to 6197)
  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"; doi: 
            "10.1016/j.cell.2023.12.032"; journalName: "Cell"; date: "2024-02-1-
            1"; volume: "187"; issue: "3"; pages: "692-711.e26"
COMMENT     Sequence Label: pAAV_FKBP-V5-TWIST1
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   826..955
                     /label=AAV2 ITR
     primer_bind     complement(1011..1030)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(1943..1945,1946..2266)
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             2282..2323
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     repeat_region   3302..3431
                     /label=AAV2 ITR
     rep_origin      3802..4315
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3889..3908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4099..4120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4797..4816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4916..4938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4976..4994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5062..5166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5167..5235,5236..6027)
                     /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"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      961 ttaatgatta acccgccatg ctacttatct acgtagccat gctctagcgg cctcggcctc
     1021 tgcataaata aaaaaaatta gtcagccatg agcttggccc attgcatacg ttgtatccat
     1081 atcataatat gtacatttat attggctcat gtccaacatt accgccatgt tgacattgat
     1141 tattgactag acggacctag agggctcttg ggcgagatga gacatcaccc actgtgtaga
     1201 agctgttgcc attgctgctg tcacagccac tccggatggg gctgccaccg cggccaggac
     1261 agtctcctcc gaccgcttcc tgggctgcgc tagggttcgg gggcgctgcc cgcacgctcc
     1321 ggcggggaag gaaatcgccc cgcgcccgcc ggaggaaggc gacggggagg gaagggggag
     1381 ggcggctagg aggcgggtgg aggggccggc cgcccgggcc aggtcgtttt tgaatggttt
     1441 gggaggacga attgttagac cccgaggaag ggaggtggga cgggggaggg ggactggaaa
     1501 gcggaaactt tcctataaaa cttcgaaaag tccctcctcc tcacgtcagg ccaatgacac
     1561 tgctgccccc aaactttccg cctgcacgga ggtataagag cctccaagtc tgcagctctc
     1621 gcccaactcc cagacacctc gcgggctctg cagcaccggc accgtttcca ggaggcctgg
     1681 cggggtgtgc gtccagccgt tgggcgcttt ctttttggac ctcggggcca tccacaccgt
     1741 cccctccccc tcccgcctcc ctccccgcct cccccgcgcg ccctccccgc ggaggtccct
     1801 cccgtccgtc ctcctgctct ctcctccgcg ggccgcatcg cccgggccgg cgccgcgcgc
     1861 gggggaagct ggcgggctga ggcgccccgc tcttctcctc tgccccgggc ccgcgaggcc
     1921 acgcgtcgcc gctcgagaga tgatgggagt gcaggtggaa accatctccc caggagacgg
     1981 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     2041 tggaaagaaa gttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     2101 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     2161 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     2221 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaatctg gatccggttc
     2281 cggcaagccc atccccaacc ccctgctggg cctggacagc acctccggtt caggctctgg
     2341 atccggttcc caggacgtgt ccagctcgcc agtctcgccg gccgacgaca gcctgagcaa
     2401 cagcgaggaa gagccagacc ggcagcagcc gccgagcggc aagcgcgggg gacgcaagcg
     2461 gcgcagcagc aggcgcagcg cgggcggcgg cgcggggccc ggcggagccg cgggtggggg
     2521 cgtcggaggc ggcgacgagc cgggcagccc ggcccagggc aagcgcggca agaagtctgc
     2581 gggctgtggc ggcggcggcg gcgcgggcgg cggcggcggc agcagcagcg gcggcgggag
     2641 tccgcagtct tacgaggagc tgcagacgca gcgggtcatg gccaacgtgc gggagcgcca
     2701 gcgcacccag tcgctgaacg aggcgttcgc cgcgctgcgg aagatcatcc ccacgctgcc
     2761 ctcggacaag ctgagcaaga ttcagaccct caagctggcg gccaggtaca tcgacttcct
     2821 ctaccaggtc ctccagagcg acgagctgga ctccaagatg gcaagctgca gctatgtggc
     2881 tcacgagcgg ctcagctacg ccttctcggt ctggaggatg gagggggcct ggtccatgtc
     2941 cgcgtcccac tagcaggcgg agccccccac cccctcagca gggccggaga cctaggtaag
     3001 gaccgcgccg ctgcacccct tcgcctctca ggtggcagac ggcaggccgg ccaggccgcg
     3061 gttcccagtc cacctcgatt tcctcccctc tcccactctc cgctcagcct tcccacctca
     3121 cttggcaccg ttgcctcgcg cccccagcgt ccccggaagg ccggtctgac cccgctaggg
     3181 agagcagtct ccagggggat gcgccctggt gaggggtgtg tgtgcgcgtg agtgtgcgtg
     3241 acaggagggc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     3301 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3361 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3421 cgagcgcgca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3481 cgcagcctga atggcgaatg gcgattccgt tgcaatggct ggcggtaata ttgttctgga
     3541 tattaccagc aaggccgata gtttgagttc ttctactcag gcaagtgatg ttattactaa
     3601 tcaaagaagt attgcgacaa cggttaattt gcgtgatgga cagactcttt tactcggtgg
     3661 cctcactgat tataaaaaca cttctcagga ttctggcgta ccgttcctgt ctaaaatccc
     3721 tttaatcggc ctcctgttta gctcccgctc tgattctaac gaggaaagca cgttatacgt
     3781 gctcgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3841 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3901 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3961 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4021 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4081 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4141 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4201 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttaaa
     4261 tatttgctta tacaatcttc ctgtttttgg ggcttttctg attatcaacc ggggtacata
     4321 tgattgacat gctagtttta cgattaccgt tcatcgattc tcttgtttgc tccagactct
     4381 caggcaatga cctgatagcc tttgtagaga cctctcaaaa atagctaccc tctccggcat
     4441 gaatttatca gctagaacgg ttgaatatca tattgatggt gatttgactg tctccggcct
     4501 ttctcacccg tttgaatctt tacctacaca ttactcaggc attgcattta aaatatatga
     4561 gggttctaaa aatttttatc cttgcgttga aataaaggct tctcccgcaa aagtattaca
     4621 gggtcataat gtttttggta caaccgattt agctttatgc tctgaggctt tattgcttaa
     4681 ttttgctaat tctttgcctt gcctgtatga tttattggat gttggaatcg cctgatgcgg
     4741 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     4801 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4861 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4921 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4981 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5041 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5101 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5161 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5281 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5341 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5401 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5461 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5581 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5701 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5761 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5881 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5941 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6001 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6061 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6121 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6181 aagatcaaag gatcttc
//
