LOCUS       sequence-349083-        4646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector mediating neuron-specific expression of rtTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4646)
  AUTHORS   Li E, Guo J, Oh SJ, Luo Y, Oliveros HC, Du W, Arano R, Kim Y, Chen 
            YT, Eitson J, Lin DT, Li Y, Roberts T, Schoggins JW, Xu W
  TITLE     Anterograde transneuronal tracing and genetic control with 
            engineered yellow fever vaccine YFV-17D.
  JOURNAL   Nat Methods. 2021 Nov 25. pii: 10.1038/s41592-021-01319-9. doi: 
            10.1038/s41592-021-01319-9.
  PUBMED    34824475
REFERENCE   2  (bases 1 to 4646)
  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: "Nat 
            Methods. 2021 Nov 25. pii: 10.1038/s41592-021-01319-9. doi: 
            10.1038/s41592-021-01319-9."
COMMENT     Sequence Label: pAAV-rtTA
FEATURES             Location/Qualifiers
     source          1..4646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        114..561
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      564..573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             595..1302
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     polyA_signal    1339..1815
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1538..1557)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   1855..1995
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1855..1984
                     /label=AAV2 ITR
     rep_origin      2070..2525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2157..2176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2367..2388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2542..2561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2661..2683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2721..2739)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2807..2911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2912..2980,2981..3772)
                     /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(3130..3149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3943..4531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4432..4451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4593..4646,1..76)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
       61 atcactaggg gttcctgcgg ccgcacgcgc gtctgcagag ggccctgcgt atgagtgcaa
      121 gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac
      181 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
      241 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
      301 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
      361 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
      421 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
      481 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
      541 gtgtcgtgcc tgagagcgca gtcgccacca tgggatccga ttgaattcgc caccatgtct
      601 aggctggaca agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc
      661 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
      721 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg
      781 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
      841 gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
      901 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
      961 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
     1021 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
     1081 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc
     1141 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
     1201 tgcgaaagcg gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc
     1261 gatgcccttg acgattttga ccttgacatg ctccccgggt aaaagcttgc ctcgagcagc
     1321 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     1381 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     1441 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     1501 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     1561 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     1621 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     1681 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     1741 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     1801 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     1861 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     1921 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     1981 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2041 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     2101 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     2161 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2221 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2281 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2341 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2401 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     2461 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     2521 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     2581 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2641 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2701 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2761 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2821 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2881 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2941 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3001 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3061 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3121 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3181 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3241 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3301 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3361 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3421 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3481 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3541 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3601 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3661 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3721 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3781 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3841 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4021 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4081 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4261 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4501 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4561 cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct
     4621 cactgaggcc gcccgggcgt cgggcg
//
