LOCUS       sequence-383801-        6985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for knocking in a C-terminal tag (dTAG-mNeonGreen-V5) for
            human SOX9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6985)
  AUTHORS   Naqvi S, Kim S, Hoskens H, Matthews HS, Spritz RA, Klein OD, 
            Hallgrimsson B, Swigut T, Claes P, Pritchard JK, Wysocka J
  TITLE     Precise modulation of transcription factor levels identifies 
            features underlying dosage sensitivity.
  JOURNAL   Nat Genet. 2023 May;55(5):841-851. doi: 10.1038/s41588-023-01366-2. 
            Epub 2023 Apr 6.
  PUBMED    37024583
REFERENCE   2  (bases 1 to 6985)
  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: 
            "10.1038/s41588-023-01366-2"; journalName: "Nat Genet"; date: 
            "2023-05"; volume: "55"; issue: "5"; pages: "841-851"
COMMENT     Sequence Label: pAAV-hSOX9-dTAG-mNeonGreen-V5
FEATURES             Location/Qualifiers
     source          1..6985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..133)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      927..1515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1416..1435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1752..1881
                     /label=AAV2 ITR
     primer_bind     complement(1937..1956)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2996..3316
                     /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="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3332..4039
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3587..3596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4061..4102
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     repeat_region   5016..5145
                     /label=AAV2 ITR
     rep_origin      5516..6029
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5603..5622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5813..5834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6511..6530)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6630..6652
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6690..6708)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6776..6880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6881..6949,6950..6985,1..756)
                     /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 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
       61 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      121 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      181 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      241 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      301 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      361 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      421 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
      481 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      541 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      601 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      661 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      721 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
      781 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
      841 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
      901 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
      961 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1021 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1081 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1141 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1201 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1261 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1321 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1381 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1441 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1501 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1561 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1621 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1681 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     1741 cattaatgca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     1801 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     1861 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
     1921 agccatgctc tagcggcctc ggcctctgca taaataaaaa aaattagtca gccatgagct
     1981 tggcccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     2041 aacattaccg ccatgttgac attgattatt gagcacagcc cttgttgatt ttctcgtgct
     2101 tgttctttta ttgtccacag ggcaatccca gggcccaccg accccaccca ccacccccaa
     2161 aaccgacgtg cagccgggca aggctgacct gaagcgagag gggcgcccct tgccagaggg
     2221 gggcagacag ccccctatcg acttccgcga cgtggacatc ggcgagctga gcagcgacgt
     2281 catctccaac atcgagacct tcgatgtcaa cgagtttgac cagtacctgc cgcccaacgg
     2341 ccacccgggg gtgccggcca cgcacggcca ggtcacctac acgggcagct acggcatcag
     2401 cagcaccgcg gccaccccgg cgagcgcggg ccacgtgtgg atgtccaagc agcaggcgcc
     2461 gccgccaccc ccgcagcagc ccccacaggc cccgccggcc ccgcaggcgc ccccgcagcc
     2521 gcaggcggcg cccccacagc agccggcggc acccccgcag cagccacagg cgcacacgct
     2581 gaccacgctg agcagcgagc cgggccagtc ccagcgaacg cacatcaaga cggagcagct
     2641 gagccccagc cactacagcg agcagcagca gcactcgccc caacagatcg cctacagccc
     2701 cttcaacctc ccacactaca gcccctccta cccgcccatc acccgctcac agtacgacta
     2761 caccgaccac cagaactcca gctcctacta cagccacgcg gcaggccagg gcaccggcct
     2821 ctactccacc ttcacctaca tgaaccccgc tcagcgcccc atgtacaccc ccatcgccga
     2881 cacctctggg gtcccttcca tcccgcagac ccacagcccc cagcactggg aacaacccgt
     2941 ctacacacag ctcactcgac ctgccggtgc aggcgctgga gccggtgcca ccggtggagt
     3001 gcaggtggaa accatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg
     3061 cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa gttgattcct cccgggacag
     3121 aaacaagccc tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg
     3181 ggttgcccag atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta
     3241 tggtgccact gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga
     3301 gcttctaaaa ctggaaggca gtggaggatc catggtgagc aagggcgagg aggataacat
     3361 ggcctctctc ccagcgacac atgagttaca catctttggc tccatcaacg gtgtggactt
     3421 tgacatggtg ggtcagggca ccggcaatcc aaatgatggt tatgaggagt taaacctgaa
     3481 gtccaccaag ggtgacctcc agttctcccc ctggattctg gtccctcata tcgggtatgg
     3541 cttccatcag tacctgccct accctgacgg gatgtcgcct ttccaggccg ccatggtaga
     3601 tggctccgga taccaagtcc atcgcacaat gcagtttgaa gatggtgcct cccttactgt
     3661 taactaccgc tacacctacg agggaagcca catcaaagga gaggcccagg tgaaggggac
     3721 tggtttccct gctgacggtc ctgtaatgac caactcgctg accgctgcgg actggtgcag
     3781 gtcgaagaag acttacccca acgacaaaac catcatcagt acctttaagt ggagttacac
     3841 cactggaaat ggcaagcgct accggagcac tgcgcggacc acctacacct ttgccaagcc
     3901 aatggcggct aactatctga agaaccagcc gatgtacgtg ttccgtaaga cggagctcaa
     3961 gcactccaag accgagctca acttcaagga gtggcaaaag gcctttaccg atgtgatggg
     4021 catggacgag ctgtacaagc tcgaggctgg agccggtgcc ggcaagccca tccccaaccc
     4081 cctgctgggc ctggacagca cctgaggagg cctcccacga agggcgaaga tggccgagat
     4141 gatcctaaaa ataaccgaag aaagagagga ccaaccagaa ttccctttgg acatttgtgt
     4201 ttttttgttt ttttattttg ttttgttttt tcttcttctt cttcttcctt aaagacattt
     4261 aagctaaagg caactcgtac ccaaatttcc aagacacaaa catgacctat ccaagcgcat
     4321 tacccacttg tggccaatca gtggccaggc caaccttggc taaatggagc agcgaaatca
     4381 acgagaaact ggacttttta aaccctcttc agagcaagcg tggaggatga tggagaatcg
     4441 tgtgatcagt gtgctaaatc tctctgcctg tttggacttt gtaattattt ttttagcagt
     4501 aattaaagaa aaaagtcctc tgtgaggaat attctctatt ttaaatattt ttagtatgta
     4561 ctgtgtatga ttcattacca ttttgagggg atttatacat atttttagat aaaattaaat
     4621 gctcttattt ttccaacagc taaactactc ttagttgaac agtgtgccct agcttttctt
     4681 gcaaccagag tatttttgta cagatttgct ttctcttaca aaaagaaaaa aaaaatcctg
     4741 ttgtattaac atttaaaaac agaattgtgt tatgtgatca gttttggggg ttaactttgc
     4801 ttaattcctc aggctttgcg atttaaggag gagctgcctt aaaaaaaaat aaaggcctta
     4861 ttttgcaatt atgggagtaa acaatagtct agagaagcat ttggtaagct ttatcatata
     4921 tatatttttt aaagaagaga aaaacacctt gagccttaaa acggtgcagc atggctacgt
     4981 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     5041 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5101 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctggc gtaatagcga
     5161 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgatt
     5221 ccgttgcaat ggctggcggt aatattgttc tggatattac cagcaaggcc gatagtttga
     5281 gttcttctac tcaggcaagt gatgttatta ctaatcaaag aagtattgcg acaacggtta
     5341 atttgcgtga tggacagact cttttactcg gtggcctcac tgattataaa aacacttctc
     5401 aggattctgg cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg tttagctccc
     5461 gctctgattc taacgaggaa agcacgttat acgtgctcgt caaagcaacc atagtacgcg
     5521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5941 aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat cttcctgttt
     6001 ttggggcttt tctgattatc aaccggggta catatgattg acatgctagt tttacgatta
     6061 ccgttcatcg attctcttgt ttgctccaga ctctcaggca atgacctgat agcctttgta
     6121 gagacctctc aaaaatagct accctctccg gcatgaattt atcagctaga acggttgaat
     6181 atcatattga tggtgatttg actgtctccg gcctttctca cccgtttgaa tctttaccta
     6241 cacattactc aggcattgca tttaaaatat atgagggttc taaaaatttt tatccttgcg
     6301 ttgaaataaa ggcttctccc gcaaaagtat tacagggtca taatgttttt ggtacaaccg
     6361 atttagcttt atgctctgag gctttattgc ttaattttgc taattctttg ccttgcctgt
     6421 atgatttatt ggatgttgga atcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6481 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6541 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     6601 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     6661 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     6721 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     6781 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6841 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6901 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6961 gctggtgaaa gtaaaagatg ctgaa
//
