LOCUS       sequence-425820-        6203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for knocking in an N-terminal tag (FKBP12F36V and V5) for
            human ALX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6203)
  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 6203)
  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-ALX1
FEATURES             Location/Qualifiers
     source          1..6203
                     /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(1989..1991,1992..2312)
                     /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             2328..2369
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     repeat_region   3308..3437
                     /label=AAV2 ITR
     rep_origin      3808..4321
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4105..4126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4803..4822)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4922..4944
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4982..5000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5068..5172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5173..5241,5242..6033)
                     /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     complement(5391..5410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
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 gcttctctct cccacccatc gccaccgcgt ccacactccg ccgcattcca
     1201 gagaaaaaga aaaggccacg gcctcgtaaa gctcccagct ggcccggacc ccgagctcct
     1261 cccggttaga agccgaagcg gcccctctac tacggcggct gcgattaaca gttgccaatg
     1321 catctggttc cttgggcttc ggattaatgt gaatcttcca cgtaaacgat ttacggatga
     1381 aataaaagcc ttttagagca atatttcctt tcaagttctt aagacgcctc taaatctttc
     1441 tggtagtgag tggccggcat gggggcgggg tggggacggg tggctcagga aggcgcaggc
     1501 agtagggagt ggaggtttaa ctgctttgcg atctcaccaa gatcttccga aatgccagtc
     1561 cactgtcacg gcggcaatga atctgggtgt caggggctcg tctctcctgg catttgtctg
     1621 aaatggagct gtctgtctgt ctatcctagg cctggacgcc ctctgcccgg gtaagtgggg
     1681 aaaatggggg tggggtaggg gcggaggggg acctggggtg gagttagtcg cccgcctatt
     1741 ttcatattca ttagaactgg gaggctgccc gaggaggcca gctgagtttc aagatataaa
     1801 agcaacttcg ggtgctcctt ttccagccgg gacgtattaa agggaccgga gcggggcagt
     1861 tcccgacgcc gcagcgctcg ctttcccttc cctcctctct ggccctccct cctcccaccc
     1921 actggctccc tcccccagcg ctctccagtt tctgtgcccc aggagctacg cgacagtctt
     1981 ccaggattat gggagtgcag gtggaaacca tctccccagg agacgggcgc accttcccca
     2041 agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaagttg
     2101 attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc
     2161 gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat
     2221 ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca catgccactc
     2281 tcgtcttcga tgtggagctt ctaaaactgg aatctggatc cggttccggc aagcccatcc
     2341 ccaaccccct gctgggcctg gacagcacct ccggttcagg ctctggatcc ggttccgagt
     2401 ttctgagcga gaagtttgcc ctcaagagcc ctccgagtaa aaacagtgac ttttacatgg
     2461 gcgcaggagg tcctctggag cacgttatgg agacgctgga caatgagtcc ttttacagca
     2521 aagcgtctgc aggcaaatgc gtgcaggcct tcggacccct gccccgcgcc gagcatcacg
     2581 tgcgcttgga gaggacctcg ccctgtcagg acagcagcgg tgagtcgcta gcgccccagc
     2641 cggagccgcc gcagcccttc cgcaatcgaa aatgcaggat cggtctgatc aggcagggag
     2701 ggagaaagga gaaaagtggg agcgagggac ctggaaggtg gaggaaggta gtggaaggtg
     2761 ctcgctttat gaagcggtgt tactgctgct gcagcctttg ggggcggagg gcggagggag
     2821 ggagagatcc agcaccgggt tgcctctgag cgattctcct ttcgctgtag ggtctggtcg
     2881 cctacggatc cctgagtcct tctgtaatat tggttcctgg agtaaactga tagggcgcta
     2941 aatagagcgt aattgagtca agacagaacg cttcctctgc gagcccgcct tgcagtccgg
     3001 gcggtaggaa ctgaaatcga tcacagctcg ttagagtacg tcactgtcgt cacgagtagg
     3061 gaaaaaaaaa ccggaaaaca aaaacaaaaa acaaaaaacc ctttgagctc ctttaggtaa
     3121 agataaatct ttttaatttt ccaagacgtg gcgatatatt ttcaaagttt gaggatccct
     3181 agatttctag cgtgtgtgaa tttgtaattt ctgctttttt ttgcgtttgc aatacaacac
     3241 gcgttgaatg tttgtctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     3301 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3361 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3421 agcgagcgag cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3481 cagttgcgca gcctgaatgg cgaatggcga ttccgttgca atggctggcg gtaatattgt
     3541 tctggatatt accagcaagg ccgatagttt gagttcttct actcaggcaa gtgatgttat
     3601 tactaatcaa agaagtattg cgacaacggt taatttgcgt gatggacaga ctcttttact
     3661 cggtggcctc actgattata aaaacacttc tcaggattct ggcgtaccgt tcctgtctaa
     3721 aatcccttta atcggcctcc tgtttagctc ccgctctgat tctaacgagg aaagcacgtt
     3781 atacgtgctc gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4261 tttaaatatt tgcttataca atcttcctgt ttttggggct tttctgatta tcaaccgggg
     4321 tacatatgat tgacatgcta gttttacgat taccgttcat cgattctctt gtttgctcca
     4381 gactctcagg caatgacctg atagcctttg tagagacctc tcaaaaatag ctaccctctc
     4441 cggcatgaat ttatcagcta gaacggttga atatcatatt gatggtgatt tgactgtctc
     4501 cggcctttct cacccgtttg aatctttacc tacacattac tcaggcattg catttaaaat
     4561 atatgagggt tctaaaaatt tttatccttg cgttgaaata aaggcttctc ccgcaaaagt
     4621 attacagggt cataatgttt ttggtacaac cgatttagct ttatgctctg aggctttatt
     4681 gcttaatttt gctaattctt tgccttgcct gtatgattta ttggatgttg gaatcgcctg
     4741 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     4801 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4861 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4921 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4981 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5041 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5101 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5161 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5221 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5281 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5341 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5401 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5461 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5521 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5581 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5641 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5701 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5761 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5821 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5881 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5941 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6001 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6061 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6121 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6181 gtagaaaaga tcaaaggatc ttc
//
