LOCUS       sequence-346765-        5592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector mediating Cre-depenent expression of NS1 gene of YFV-17D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5592)
  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 5592)
  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-DIO-NS1
FEATURES             Location/Qualifiers
     source          1..5592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   76..216
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   87..216
                     /label=AAV2 ITR
     polyA_signal    239..446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     435..452
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    487..1075
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             593..604
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1002..1022
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(1099..1132)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1143..1176)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2348..2381
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2392..2425
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(2458..2905)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   2941..3070
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3132..3720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3212..3231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3891..4682,4683..4751))
                     /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     4514..4533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4752..4856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4924..4942
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4980..5002)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5102..5121
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5138..5592,1))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5275..5296)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5487..5506
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
       61 accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
      121 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      181 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgtc
      241 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
      301 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
      361 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
      421 agatggctgg caactagaag gcacagtcga ggcagatcga tctctcgagc agcgctcggt
      481 atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga aggcgaagac
      541 gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg
      601 ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca taggcgagca
      661 gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg cccaacagcc
      721 gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaataggga
      781 gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc caaccagtgg
      841 gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg ggccacaact
      901 cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa gccatacggg
      961 aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa
     1021 catagttaag aataccagtc aatctttcac aaattttgta atccagaggt tgattatcga
     1081 taagcttgat gatcttcgat aacttcgtat aggatacttt atacgaagtt attgcgattc
     1141 ggataacttc gtatagcata cattatacga agttatcaag gccggccagg ttggcgcgcc
     1201 accatgaaca tgacaatgtc catgagcatg atcttggtag gagtgatcat gatgtttttg
     1261 tctctaggag ttggggcgga tcaaggatgc gccatcaact ttggcaagag agagctcaag
     1321 tgcggagatg gtatcttcat atttagagac tctgatgact ggctgaacaa gtactcatac
     1381 tatccagaag atcctgtgaa gcttgcatca atagtgaaag cctcttttga agaagggaag
     1441 tgtggcctaa attcagttga ctcccttgag catgagatgt ggagaagcag ggcagatgag
     1501 atcaatgcca tttttgagga aaacgaggtg gacatttctg ttgtcgtgca ggatccaaag
     1561 aatgtttacc agagaggaac tcatccattt tccagaattc gggatggtct gcagtatggt
     1621 tggaagactt ggggtaagaa ccttgtgttc tccccaggga ggaagaatgg aagcttcatc
     1681 atagatggaa agtccaggaa agaatgcccg ttttcaaacc gggtctggaa ttctttccag
     1741 atagaggagt ttgggacggg agtgttcacc acacgcgtgt acatggacgc agtctttgaa
     1801 tacaccatag actgcgatgg atctatcttg ggtgcagcgg tgaacggaaa aaagagtgcc
     1861 catggctctc caacattttg gatgggaagt catgaagtaa atgggacatg gatgatccac
     1921 accttggagg cattagatta caaggagtgt gagtggccac tgacacatac gattggaaca
     1981 tcagttgaag agagtgaaat gttcatgccg agatcaatcg gaggcccagt tagctctcac
     2041 aatcatatcc ctggatacaa ggttcagacg aacggacctt ggatgcaggt accactagaa
     2101 gtgaagagag aagcttgccc agggactagc gtgatcattg atggcaactg tgatggacgg
     2161 ggaaaatcaa ccagatccac cacggatagc gggaaagtta ttcctgaatg gtgttgccgc
     2221 tcctgcacaa tgccgcctgt gagcttccat ggtagtgatg ggtgttggta tcccatggaa
     2281 attaggccaa ggaaaacgca tgaaagccat ctggtgcgct cctgggttac agcttaaggc
     2341 gcgccaaata acttcgtata aagtatccta tacgaagtta ttgcgattcg gataacttcg
     2401 tataatgtat gctatacgaa gttatcgacg tcgacctgaa gggtaccgtc cacgcgtctg
     2461 cgctctcagg cacgacacga ctcctccgct gcccaccgca gactgaggca gcgctgagtc
     2521 gccggcgccg cagcgcagat ggtcgcgccc gtgcccccct atctcgcgcc tcgcgtggtg
     2581 cggtccggct gggccggcgg cggcgcggac gcgaccaagg tggccgggaa ggggagtttg
     2641 cgggggaccg gcgagtgacg tcagcgcgcc ttcagtgctg aggcggcggt ggcgcgcgcc
     2701 gccaggcggg ggcgaaggca ctgtccgcgg tgctgaagct ggcagtgcgc acgcgcctcg
     2761 ccgcatcctg tttcccctcc ccctctctga taggggatgc gcaatttggg gaatgggggt
     2821 tgggtgcttg tccagtgggt cggggtcggt cgtcaggtag gcacccccac cccgcctcat
     2881 cctggtccta aaacccactt gcactcatac gcagggccct ctgcagacgc gtgcggccgc
     2941 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3001 ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3061 tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3121 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3181 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3241 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3301 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3361 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3421 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3481 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3541 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3601 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3661 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3781 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4021 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4861 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4921 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4981 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     5041 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     5101 gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg
     5161 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc
     5221 ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt
     5281 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5341 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
     5401 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5461 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg
     5521 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5581 ctacagggcg cg
//
