LOCUS       sequence-234911-        7460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector, pTet bidi (bidirectional tetracycline-responsive 
            promoter) driving Tet-dependent rtTA and Luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7460)
  AUTHORS   Chtarto A, Humbert-Claude M, Bockstael O, Das AT, Boutry S, Breger 
            LS, Klaver B, Melas C, Barroso-Chinea P, Gonzalez-Hernandez T, 
            Muller RN, DeWitte O, Levivier M, Lundberg C, Berkhout B, Tenenbaum 
            L
  TITLE     A regulatable AAV vector mediating GDNF biological effects at 
            clinically-approved sub-antimicrobial doxycycline doses.
  JOURNAL   Mol Ther Methods Clin Dev. 2016 Mar 30;5:16027. doi: 
            10.1038/mtm.2016.27. eCollection 2016.
  PUBMED    27069954
REFERENCE   2  (bases 1 to 7460)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2016-03-30"; volume: "5:16027. doi"; 
            pages: " 10.1038/mtm.2016.27. eCollection 2016"
COMMENT     Sequence Label: pAC1-V16-Luc
FEATURES             Location/Qualifiers
     source          1..7460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   41..170
                     /label=AAV2 ITR
     polyA_signal    517..651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(661..1407)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKIINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(1509..1547)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(1580..1598,1599..1621,1622..1640,1641..1663,1664..1682,
                     1683..1705,1706..1724,1725..1747,1748..1766,1767..1789,
                     1790..1808,1809..1831,1832..1850)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(1622..1640,1641..1663,1664..1682,1683..1705,1706..1724,
                     1725..1747,1748..1766,1767..1789,1790..1808,1809..1831,
                     1832..1850,1851..1873,1874..1892)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     CDS             2032..3684
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2098..2116)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          3783..3848
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3978..3998
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4423..4557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4846..5434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4926..4945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5605..6396,6397..6465))
                     /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     6228..6247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6466..6570)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6638..6656
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6694..6716)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6816..6835
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6902..7357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7039..7060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7251..7270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 krrsgggggg gggggggggg gggggggggc cactccctct ctgcgcgctc gctcgctcac
       61 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
      121 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct cagatcttca
      181 gcttggtcga gctggatact tcccgtccgc cagggggaca tgccggcgat gctgaaggtc
      241 gcgcgcattc ccgatgaaga ggccggttac cgcctgttga cctggtggga cgggcagggc
      301 gccgcccgag tcttcgcctc ggcggcgggc gctctgctca tggagcgcgc gtccggggcc
      361 ggggaccttg cacagatagc gtggtccggc caggacgacg aggcttgcag gatcataatc
      421 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
      481 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
      541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
      601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccttacttag
      661 ttacccgggg agcatgtcaa ggtcaaaatc gtcaagagcg tcagcaggca gcatatcaag
      721 gtcaaagtcg tcaagggcat cggctgggag catgtctaag tcaaaatcgt caagggcgtc
      781 ggtcggcccg ccgctttcgc actttagctg tttctccagg ccacatatga ttagttccag
      841 gccgaaaagg aaggcaggtt cggctccctg ccggtcgaac agctcaattg cttgtttcag
      901 aagtgggggc atagaatcgg tggtaggtgt ctctctttcc tcttttgcta cttgatgctc
      961 ctgttcctcc aatacgcagc ccagtgtaaa gtggcccacg gcggacagag cgtacagtgc
     1021 gttctccagg gagaagcctt gctgacacag gaacgcgagc tgattttcca gggtttcgta
     1081 ctgtttctct gttgggcggg tgccgagatg cactttagcc ccgtcgcgat gtgagaggag
     1141 agcacagcgg tatgacttgg cgttgttccg cagaaagtct tgccatgact cgccttccag
     1201 ggggcaggag tgggtatgat gcctgtccag catctcgatt ggcagggcat cgagcagggc
     1261 ccgcttgttc ttcacgtgcc agtacagggt aggctgctca actcccagct tttgagcgag
     1321 tttccttgtc gtcaggcctt cgataccgac tccattgagt aattccagag cagagtttat
     1381 gattttgctc ttgtccagtc tagacatatg aattcggggc cgcggaggct ggatcggtcc
     1441 cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc
     1501 actaaacgag ctctgcttat ataggcctcc caccgtacac gcctacctcg acccgggtac
     1561 cgagctcgac tttcactttt ctctatcact gatagggagt ggtaaactcg actttcactt
     1621 ttctctatca ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg
     1681 gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgactt
     1741 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
     1801 gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact
     1861 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgacctat ataagcagag
     1921 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1981 gaagacaccg ggaccgatcc agcctccgcg gccccgaatt ctgcagtcga catggaagac
     2041 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     2101 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     2161 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     2221 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     2281 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     2341 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     2401 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     2461 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     2521 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     2581 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     2641 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     2701 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     2761 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     2821 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     2881 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     2941 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     3001 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     3061 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     3121 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     3181 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     3241 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     3301 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     3361 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     3421 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     3481 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     3541 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     3601 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     3661 aagggcggaa agatcgccgt gtaagcggcc gcgactctag ctagaggatc tttgtgaagg
     3721 aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta
     3781 aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     3841 tattttagat tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg
     3901 aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact
     3961 ctcaacattc tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt
     4021 cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg
     4081 ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt
     4141 ctgtaacctt tataagtagg cataacagtt ataatcataa catactgttt tttcttactc
     4201 cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct
     4261 ttttaatttg taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc
     4321 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4381 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgaa
     4561 gatctgagga acccctagtg atggagttgg ccactcccyc yctgcagcct gcattaatga
     4621 atcggccaac gcgcggggag aggcggtttg cgymwykssc kctctkcgcg ctcsctcgct
     4681 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4741 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4801 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4861 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5041 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5161 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5281 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5341 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5401 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5461 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5521 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5581 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5641 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5701 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5761 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5821 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5881 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5941 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6001 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6061 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6121 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6181 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6241 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6301 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6361 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6421 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6481 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6541 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6601 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6661 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6721 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6781 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6841 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6901 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     6961 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     7021 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     7081 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     7141 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     7201 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     7261 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     7321 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     7381 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccaggct gcaggggggg
     7441 gggggggggg gccccccccs
//
