LOCUS       Exported                6496 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector, pTet bidi (bidirectional tetracycline-responsive 
            promoter) driving Tet-dependent rtTA and hGDNF expression.
ACCESSION   .
VERSION     .
KEYWORDS    pAC1-V16-hGDNF
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  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 6496)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6496
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    1..271
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    43..313
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     intron          1280..1345
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1475..1495
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1920..2054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2171..2188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2342..2930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2422..2441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3101..3961)
                     /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     3724..3743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3962..4066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4134..4152
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4190..4212)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4312..4331
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4398..4853)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4535..4556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4747..4766
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4958..5087
                     /label=AAV2 ITR
     polyA_signal    5434..5568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5578..6324)
                     /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(6426..6464)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc actgataggg
       61 agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt
      121 cacttttctc tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg
      181 atagggagtg gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc
      241 gactttcact tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta
      301 tcactgatag ggagtggtaa actcgaccta tataagcaga gctcgtttag tgaaccgtca
      361 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      421 cagcctccgc ggccccgaat tctgcagtcg acggtaccgc gggcccggga tccactagtt
      481 ctagcctcga gaattagctt cgccaccatg aagttatggg atgtcgtggc tgtctgcctg
      541 gtgctgctcc acaccgcgtc cgccttcccg ctgcccgccg gtaagaggcc tcccgaggcg
      601 cccgccgaag accgctccct cggccgccgc cgcgcgccct tcgcgctgag cagtgactca
      661 aatatgccag aggattatcc tgatcagttc gatgatgtca tggattttat tcaagccacc
      721 attaaaagac tgaaaaggtc accagataaa caaatggcag tgcttcctag aagagagcgg
      781 aatcggcagg ctgcagctgc caacccagag aattccagag gaaaaggtcg gagaggccag
      841 aggggcaaaa accggggttg tgtcttaact gcaatacatt taaatgtcac tgacttgggt
      901 ctgggctatg aaaccaagga ggaactgatt tttaggtact gcagcggctc ttgcgatgca
      961 gctgagacaa cgtacgacaa aatattgaaa aacttatcca gaaatagaag gctggtgagt
     1021 gacaaagtag ggcaggcatg ttgcagaccc atcgcctttg atgatgacct gtcgttttta
     1081 gatgataacc tggtttacca tattctaaga aagcattccg ctaaaaggtg tggatgtatc
     1141 tgaggatcaa ttcacgcgtg gtacctctag agtcgacccg ggcggccgcg actctagcta
     1201 gaggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac
     1261 agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact
     1321 gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg
     1381 gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga
     1441 tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga
     1501 ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag
     1561 aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa
     1621 aattatggaa aaatattctg taacctttat aagtaggcat aacagttata atcataacat
     1681 actgtttttt cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa
     1741 attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag
     1801 tgccttgact agagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1861 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1981 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2041 atcatgtctg gatcgaagat ctgaggaacc cctagtgatg gagttggcca ctccctctct
     2101 gcagcctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2161 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2221 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2281 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2341 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2401 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2461 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2521 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2581 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2641 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2701 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2761 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2821 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2941 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3001 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3061 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3121 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3181 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3241 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3301 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3361 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3421 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3481 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3541 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3601 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3661 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3721 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3781 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3841 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3901 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4021 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4081 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4141 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4201 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4261 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4321 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4381 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4441 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4501 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4561 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4621 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4681 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4741 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4801 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     4861 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4921 caggckgsrg gggggggggg ggggggccac tccctctctg cgcgctcgct cgctcactga
     4981 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     5041 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctcag atcttcagct
     5101 tggtcgagct ggatacttcc cgtccgccag ggggacatgc cggcgatgct gaaggtcgcg
     5161 cgcattcccg atgaagaggc cggttaccgc ctgttgacct ggtgggacgg gcagggcgcc
     5221 gcccgagtct tcgcctcggc ggcgggcgct ctgctcatgg agcgcgcgtc cggggccggg
     5281 gaccttgcac agatagcgtg gtccggccag gacgacgagg cttgcaggat cataatcagc
     5341 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     5401 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     5461 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5521 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcct tacttagtta
     5581 cccggggagc atgtcaaggt caaaatcgtc aagagcgtca gcaggcagca tatcaaggtc
     5641 aaagtcgtca agggcatcgg ctgggagcat gtctaagtca aaatcgtcaa gggcgtcggt
     5701 cggcccgccg ctttcgcact ttagctgttt ctccaggcca catatgatta gttccaggcc
     5761 gaaaaggaag gcaggttcgg ctccctgccg gtcgaacagc tcaattgctt gtttcagaag
     5821 tgggggcata gaatcggtgg taggtgtctc tctttcctct tttgctactt gatgctcctg
     5881 ttcctccaat acgcagccca gtgtaaagtg gcccacggcg gacagagcgt acagtgcgtt
     5941 ctccagggag aagccttgct gacacaggaa cgcgagctga ttttccaggg tttcgtactg
     6001 tttctctgtt gggcgggtgc cgagatgcac tttagccccg tcgcgatgtg agaggagagc
     6061 acagcggtat gacttggcgt tgttccgcag aaagtcttgc catgactcgc cttccagggg
     6121 gcaggagtgg gtatgatgcc tgtccagcat ctcgattggc agggcatcga gcagggcccg
     6181 cttgttcttc acgtgccagt acagggtagg ctgctcaact cccagctttt gagcgagttt
     6241 ccttgtcgtc aggccttcga taccgactcc attgagtaat tccagagcag agtttatgat
     6301 tttgctcttg tccagtctag acatatgaat tcggggccgc ggaggctgga tcggtcccgg
     6361 tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact
     6421 aaacgagctc tgcttatata ggcctcccac cgtacacgcc tacctcgacc cgggtaccga
     6481 gctcgacttt cacttt
//
