LOCUS       sequence-365743-        5082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tTA dependent gene expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5082)
  AUTHORS   Amo R, Matias S, Yamanaka A, Tanaka KF, Uchida N, Watabe-Uchida M
  TITLE     A gradual temporal shift of dopamine responses mirrors the 
            progression of temporal difference error in machine learning.
  JOURNAL   Nat Neurosci. 2022 Aug;25(8):1082-1092. doi: 
            10.1038/s41593-022-01109-2. Epub 2022 Jul 7.
  PUBMED    35798979
REFERENCE   2  (bases 1 to 5082)
  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/s41593-022-01109-2"; journalName: "Nat Neurosci"; date: 
            "2022-08"; volume: "25"; issue: "8"; pages: "1082-1092"
COMMENT     Sequence Label: pAAV-TetO(3G)-WPRE
FEATURES             Location/Qualifiers
     source          1..5082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   11..136
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    156..174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    192..210
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    228..246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    264..282
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    300..318
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    336..354
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    372..390
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     436..460
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          538..1110
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1118..1137
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1165..1184
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1165..1183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1188..1221)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1239..1827
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1292..1312)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1710..1721)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1839..2315
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2038..2057)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2355..2495
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2355..2484
                     /label=AAV2 ITR
     rep_origin      2570..3025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2657..2676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2867..2888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3042..3061)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3161..3183
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3221..3239)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3307..3411
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3412..3480,3481..4272)
                     /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(3630..3649)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4443..5031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4932..4951
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
       61 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttcctgcgg ccgcacgcgt tttactccct atcagtgata gagaacgtat
      181 gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga
      241 tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact
      301 ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat
      361 atccagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg
      421 cgcctataaa agcagagctc gtttagtgaa ccgtcagatc gcctggagca attccacaac
      481 acttttgtct tataccaact ttccgtacca cttcctaccc tcgttgagaa cttcagggtg
      541 agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat gttatatgga
      601 gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta tcaccatgga
      661 ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt gtctcctctt
      721 attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt gtaacgaatt
      781 tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc actttttttt
      841 caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac aattgttata
      901 attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg aaatattctt
      961 attggtagaa acaactacat cctggtcatc atcctgcctt tctccttatg gttacaatga
     1021 tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc ctctgctaac
     1081 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     1141 tctcatcatt ttggcaaaga attgtaatac gactcactat agggcgaata acttcgtata
     1201 gcatacatta tacgaagtta ttgtcgactg aattcgataa tcaacctctg gattacaaaa
     1261 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     1321 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     1381 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     1441 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     1501 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     1561 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1621 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     1681 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     1741 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     1801 ggatctccct ttgggccgcc tccccgcatc ggatctacgg gtggcatccc tgtgacccct
     1861 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     1921 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     1981 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     2041 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2101 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2161 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2221 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2281 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     2341 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2401 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2461 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2521 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2581 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2641 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2701 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2761 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2821 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2881 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2941 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3001 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3061 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3121 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3181 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3241 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3301 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3361 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3421 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3481 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3541 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3601 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3661 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3721 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3781 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3841 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3901 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3961 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4021 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4081 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4141 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4201 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4261 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4321 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4381 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4441 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4501 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4561 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4621 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4681 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4741 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4801 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4861 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4921 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4981 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5041 aacgcggcct ttttacggtt cctggccttt tgctggcctt tt
//
