LOCUS       sequence-349086-        6502 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV vector mediating bicistronic expression of NS1 gene of YFV-17D 
            and dTomato with NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  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 6502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Syn-NS1
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        133..580
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1741..1797
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1810..1830
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1831..2529
                     /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(1961..1979)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2229..2248
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2423..2446
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2579..3167
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2632..2652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3050..3061)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3199..3675
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3398..3417)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3715..3855
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3715..3844
                     /label=AAV2 ITR
     rep_origin      3930..4385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4017..4036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4227..4248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4402..4421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4521..4543
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4581..4599)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4667..4771
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4772..4840,4841..5632)
                     /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(4990..5009)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5803..6391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6292..6311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6453..6502,1..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 gcgtttaatt aagtgtctag actgcagagg
      121 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
      181 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
      241 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
      301 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
      361 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
      421 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
      481 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
      541 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggctagagga
      601 tccgccacca tgaacatgac aatgtccatg agcatgatct tggtaggagt gatcatgatg
      661 tttttgtctc taggagttgg ggcggatcaa ggatgcgcca tcaactttgg caagagagag
      721 ctcaagtgcg gagatggtat cttcatattt agagactctg atgactggct gaacaagtac
      781 tcatactatc cagaagatcc tgtgaagctt gcatcaatag tgaaagcctc ttttgaagaa
      841 gggaagtgtg gcctaaattc agttgactcc cttgagcatg agatgtggag aagcagggca
      901 gatgagatca atgccatttt tgaggaaaac gaggtggaca tttctgttgt cgtgcaggat
      961 ccaaagaatg tttaccagag aggaactcat ccattttcca gaattcggga tggtctgcag
     1021 tatggttgga agacttgggg taagaacctt gtgttctccc cagggaggaa gaatggaagc
     1081 ttcatcatag atggaaagtc caggaaagaa tgcccgtttt caaaccgggt ctggaattct
     1141 ttccagatag aggagtttgg gacgggagtg ttcaccacac gcgtgtacat ggacgcagtc
     1201 tttgaataca ccatagactg cgatggatct atcttgggtg cagcggtgaa cggaaaaaag
     1261 agtgcccatg gctctccaac attttggatg ggaagtcatg aagtaaatgg gacatggatg
     1321 atccacacct tggaggcatt agattacaag gagtgtgagt ggccactgac acatacgatt
     1381 ggaacatcag ttgaagagag tgaaatgttc atgccgagat caatcggagg cccagttagc
     1441 tctcacaatc atatccctgg atacaaggtt cagacgaacg gaccttggat gcaggtacca
     1501 ctagaagtga agagagaagc ttgcccaggg actagcgtga tcattgatgg caactgtgat
     1561 ggacggggaa aatcaaccag atccaccacg gatagcggga aagttattcc tgaatggtgt
     1621 tgccgctcct gcacaatgcc gcctgtgagc ttccatggta gtgatgggtg ttggtatccc
     1681 atggaaatta ggccaaggaa aacgcatgaa agccatctgg tgcgctcctg ggttacagct
     1741 gccactaact tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccatct
     1801 agaatggccc ccaagaagaa gaggaaggtg gtgagcaagg gcgaggaggt catcaaagag
     1861 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag
     1921 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1981 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag
     2041 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc
     2101 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac
     2161 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc
     2221 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     2281 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc
     2341 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc
     2401 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     2461 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag
     2521 ctgtacaagt aagtcgacgg cgcgcccctg cagggaattc gatatcaagc ttatcgataa
     2581 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2641 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2701 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2761 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2821 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2881 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2941 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3001 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3061 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3121 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     3181 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     3241 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     3301 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     3361 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     3421 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     3481 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     3541 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     3601 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     3661 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     3721 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3781 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3841 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3901 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3961 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4021 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4081 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4141 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4201 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4381 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4441 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4501 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4561 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4621 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4681 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4741 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4801 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4861 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4921 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4981 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5041 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5101 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5161 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5221 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5281 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5341 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5401 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5461 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5521 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5581 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5641 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5701 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5761 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5821 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5881 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5941 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6001 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6061 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6361 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6421 cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct
     6481 cactgaggcc gcccgggcgt cg
//
