LOCUS       sequence-349089-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector mediating neuron-specific expression of tTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4683)
  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 4683)
  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: AAV-tTA
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        118..565
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      568..577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(596..1213,1214..1219,1220..1255,1256..1258,1259..1294,
                     1295..1297,1298..1333,1334..1342)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    1380..1856
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1579..1598)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   1896..2036
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1896..2025
                     /label=AAV2 ITR
     rep_origin      2111..2566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2198..2217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2408..2429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2583..2602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2702..2724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2762..2780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2848..2952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2953..3021,3022..3813)
                     /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(3171..3190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3984..4572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4473..4492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4634..4683,1..80)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
       61 ctccatcact aggggttcct gcggccgcac gcgcgtctgc agagggccct gcgtatgagt
      121 gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga cgaccgaccc
      181 cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag
      241 ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac
      301 agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct
      361 gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc
      421 cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga
      481 ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg
      541 agtcgtgtcg tgcctgagag cgcagtcgcc accatgggat ccgattgaat tcaccatgtc
      601 tagactggac aagagcaaag tcataaactc tgctctggaa ttactcaatg aagtcggtat
      661 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
      721 ctggcacgtg aagaacaagc gggccctgct cgatgccctg gcaatcgaga tgctggacag
      781 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
      841 cgccaagtca ttccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
      901 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
      961 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     1021 gggctgcgta ttggaggatc aggagcatca agtagcaaaa gaggaaagag agacacctac
     1081 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc atcagggagc
     1141 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     1201 gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca tgctcccagc
     1261 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
     1321 ccttgacatg ctccccgggt aacaagcttg cctcgagcag cgctgctcga gagatctacg
     1381 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     1441 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     1501 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     1561 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     1621 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     1681 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     1741 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     1801 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     1861 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     1921 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     1981 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2041 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2101 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2161 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2221 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2281 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2341 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2401 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2461 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2521 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2581 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2641 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2701 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2761 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2941 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3001 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3061 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3121 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3181 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3241 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3301 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3361 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3421 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3481 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3541 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3601 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3661 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3721 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3781 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3841 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3901 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4501 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4561 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4621 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
     4681 tcg
//
