LOCUS       sequence-349702-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector mediating inducible bicistronic expression of NS1 gene of
            YFV-17D and dTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  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 5816)
  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-TRE-NS1-dTomato
FEATURES             Location/Qualifiers
     source          1..5816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..46
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    63..81
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    99..117
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    135..153
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    170..188
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    1504..1980
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1703..1722)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2020..2160
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2020..2149
                     /label=AAV2 ITR
     rep_origin      2235..2690
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2322..2341)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2532..2553
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2707..2726)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2826..2848
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2886..2904)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2972..3076
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3077..3145,3146..3937)
                     /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(3295..3314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4108..4696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4597..4616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4895..4976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4920..4939
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4994..5692)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(5077..5100)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5275..5294)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5544..5562
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(5693..5713)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        join(5727..5816,1..298)
                     /label=bidirectional TRE promoter
                     /note="Tet-responsive bidirectional promoter PTight-BI, 
                     consisting of seven tet operator sequences flanked on each 
                     side by the minimal CMV promoter"
     protein_bind    join(5808..5816,1..10)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
       61 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      121 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
      181 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
      241 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      301 gagatttcga gctcggtacc aataatcagc caccatgaac atgacaatgt ccatgagcat
      361 gatcttggta ggagtgatca tgatgttttt gtctctagga gttggggcgg atcaaggatg
      421 cgccatcaac tttggcaaga gagagctcaa gtgcggagat ggtatcttca tatttagaga
      481 ctctgatgac tggctgaaca agtactcata ctatccagaa gatcctgtga agcttgcatc
      541 aatagtgaaa gcctcttttg aagaagggaa gtgtggccta aattcagttg actcccttga
      601 gcatgagatg tggagaagca gggcagatga gatcaatgcc atttttgagg aaaacgaggt
      661 ggacatttct gttgtcgtgc aggatccaaa gaatgtttac cagagaggaa ctcatccatt
      721 ttccagaatt cgggatggtc tgcagtatgg ttggaagact tggggtaaga accttgtgtt
      781 ctccccaggg aggaagaatg gaagcttcat catagatgga aagtccagga aagaatgccc
      841 gttttcaaac cgggtctgga attctttcca gatagaggag tttgggacgg gagtgttcac
      901 cacacgcgtg tacatggacg cagtctttga atacaccata gactgcgatg gatctatctt
      961 gggtgcagcg gtgaacggaa aaaagagtgc ccatggctct ccaacatttt ggatgggaag
     1021 tcatgaagta aatgggacat ggatgatcca caccttggag gcattagatt acaaggagtg
     1081 tgagtggcca ctgacacata cgattggaac atcagttgaa gagagtgaaa tgttcatgcc
     1141 gagatcaatc ggaggcccag ttagctctca caatcatatc cctggataca aggttcagac
     1201 gaacggacct tggatgcagg taccactaga agtgaagaga gaagcttgcc cagggactag
     1261 cgtgatcatt gatggcaact gtgatggacg gggaaaatca accagatcca ccacggatag
     1321 cgggaaagtt attcctgaat ggtgttgccg ctcctgcaca atgccgcctg tgagcttcca
     1381 tggtagtgat gggtgttggt atcccatgga aattaggcca aggaaaacgc atgaaagcca
     1441 tctggtgcgc tcctgggtta cagcttgaat aaatcgatac cgagcgctgc tcgagagatc
     1501 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     1561 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     1621 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     1681 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     1741 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     1801 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     1861 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     1921 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     1981 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     2041 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2101 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     2161 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2221 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2281 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     2341 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2401 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2461 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2521 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2581 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2641 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2701 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2761 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2821 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2881 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2941 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3001 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3061 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3121 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3181 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3241 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3301 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3361 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3421 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3481 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3541 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3601 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3661 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3721 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3781 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3841 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3901 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3961 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4021 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4081 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4141 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4201 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4261 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4321 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4381 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4441 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4501 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4561 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4621 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4681 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4741 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     4801 ggcgtcgggc gacctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     4861 ctaggggttc ctgcggccgc acgcgtattt gtcggcagtg aaaaaaatgc tttatttgtg
     4921 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4981 acaacaattg ttacttgtac agctcgtcca tgccgtacag gaacaggtgg tggcggccct
     5041 cggagcgctc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccagct
     5101 tggtgtccac gtagtagtag ccgggcagtt gcacgggctt cttggccatg tagatggtct
     5161 tgaactccac caggtagtgg ccgccgtcct tcagcttcag ggcctggtgg atctcgccct
     5221 tcagcacgcc gtcgcggggg tacaggcgct cggtggaggc ctcccagccc atggtcttct
     5281 tctgcattac ggggccgtcg ggggggaagt tggtgccgcg catcttcacc ttgtagatca
     5341 gcgtgccgtc ctgcagggag gagtcctggg tcacggtcac cagaccgccg tcctcgaagt
     5401 tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcttgtaa tcggggatgt
     5461 cggcggggtg cttcacgtac gccttggagc cgtacatgaa ctggggggac aggatgtccc
     5521 aggcgaaggg cagggggccg cccttggtca ccttcagctt ggcggtctgg gtgccctcgt
     5581 aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcatg gagccctcca
     5641 tgcgcacctt gaagcgcatg aactctttga tgacctcctc gcccttgctc accaccttcc
     5701 tcttcttctt gggcatgaat tctccaggcg atctgacggt tcactaaacg agctctgctt
     5761 atataggcct cccaccgtac acgccacctc gacatactcg agtttactcc ctatca
//
