LOCUS       sequence-404980-        6227 bp    DNA     circular SYN 14-APR-2025
DEFINITION  rAAV9 cap acceptor vector for CREATE libraries.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6227)
  AUTHORS   Chuapoco MR, Flytzanis NC, Goeden N, Christopher Octeau J, Roxas KM,
            Chan KY, Scherrer J, Winchester J, Blackburn RJ, Campos LJ, Man KNM,
            Sun J, Chen X, Lefevre A, Singh VP, Arokiaraj CM, Shay TF, 
            Vendemiatti J, Jang MJ, Mich JK, Bishaw Y, Gore BB, Omstead V, 
            Taskin N, Weed N, Levi BP, Ting JT, Miller CT, Deverman BE, Pickel 
            J, Tian L, Fox AS, Gradinaru V
  TITLE     Adeno-associated viral vectors for functional intravenous gene 
            transfer throughout the non-human primate brain.
  JOURNAL   Nat Nanotechnol. 2023 Jul 10. doi: 10.1038/s41565-023-01419-x.
  PUBMED    37430038
REFERENCE   2  (bases 1 to 6227)
  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 
            Nanotechnol. 2023 Jul 10. doi: 10.1038/s41565-023-01419-x."
COMMENT     Sequence Label: rAAV9-CAP-in-cis-Lox-AAV
FEATURES             Location/Qualifiers
     source          1..6227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        130..528
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      531..540
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             537..1247
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(682..700)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     950..969
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3615..3626
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3780..3813)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3835..3956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3846..3865)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3900..3919
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    4059..4092
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   4105..4234
                     /label=AAV2 ITR
     primer_bind     complement(4299..4317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4385..4489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4490..4558,4559..5350)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4708..4727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5521..6109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6010..6029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc wwwgcccggg
       61 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      121 cctactagtg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
      181 cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc
      241 tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg
      301 acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa
      361 caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg
      421 aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg
      481 gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggcg gccgccatgg
      541 tcagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
      601 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      661 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
      721 cctgggacat cctgtcccct caattcatgt atggctccaa ggcctacgtg aagcaccccg
      781 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
      841 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctcctta caagacggcg
      901 agttcatcta caaagtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
      961 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1021 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1081 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1141 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     1201 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaagg atcccctccc
     1261 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctcc tcctaggaat
     1321 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtagatc tgttcaaatt
     1381 tgaactgact aagcggctcc cgccagattt tggcaagatt actaagcagg aagtcaagga
     1441 cttttttgct tgggcaaagg tcaatcaggt gccggtgact cacgagttta aagttcccag
     1501 ggaattggcg ggaactaaag gggcggagaa atctctaaaa cgcccactgg gtgacgtcac
     1561 caatactagc tataaaagtc tggagaagcg ggccaggctc tcatttgttc ccgagacgcc
     1621 tcgcagttca gacgtgactg ttgatcccgc tcctctgcga ccgctagctt cgatcaacta
     1681 cgcggacagg taccaaaaca aatgttctcg tcacgtgggc atgaatctga tgctgtttcc
     1741 ctgcagacaa tgcgagagac tgaatcagaa ttcaaatatc tgcttcactc acggtgtcaa
     1801 agactgttta gagtgctttc ccgtgtcaga atctcaaccc gtttctgtcg tcaaaaaggc
     1861 gtatcagaaa ctgtgctaca ttcatcacat catgggaaag gtgccagacg cttgcactgc
     1921 ttgcgacctg gtcaatgtgg acttggatga ctgtgtttct gaacaataaa tgacttaaac
     1981 caggtatggc tgccgatggt tatcttccag attggctcga ggacaacctt agtgaaggaa
     2041 ttcgcgagtg gtgggctttg aaacctggag cccctcaacc caaggcaaat caacaacatc
     2101 aagacaacgc tcgaggtctt gtgcttccgg gttacaaata ccttggaccc ggcaacggac
     2161 tcgacaaggg ggagccggtc aacgcagcag acgcggcggc cctcgagcac gacaaggcct
     2221 acgaccagca gctcaaggcc ggagacaacc cgtacctcaa gtacaaccac gccgacgccg
     2281 agttccagga gcggctcaaa gaagatacgt cttttggggg caacctcggg cgagcagtct
     2341 tccaggccaa aaagaggctt cttgaacctc ttggtctggt tgaggaagcg gctaagacgg
     2401 ctcctggaaa gaagaggcct gtagagcagt ctcctcagga accggactcc tccgcgggta
     2461 ttggcaaatc gggtgcacag cccgctaaaa agagactcaa tttcggtcag actggcgaca
     2521 cagagtcagt cccagaccct caaccaatcg gagaacctcc cgcagccccc tcaggtgtgg
     2581 gatctcttac aatggcttca ggtggtggcg caccagtggc agacaataac gaaggtgccg
     2641 atggagtggg tagttcctcg ggaaattggc attgcgattc ccaatggctg ggggacagag
     2701 tcatcaccac cagcacccga acctgggccc tgcccaccta caacaatcac ctctacaagc
     2761 aaatctccaa cagcacatct ggaggatctt caaatgacaa cgcctacttc ggctacagca
     2821 ccccctgggg gtattttgac ttcaacagat tccactgcca cttctcacca cgtgactggc
     2881 agcgactcat caacaacaac tggggattcc ggcctaagcg actcaacttc aagctcttca
     2941 acattcaggt caaagaggtt acggacaaca atggagtcaa gaccatcgcc aataacctta
     3001 ccagcacggt ccaggtcttc acggactcag actatcagct cccgtacgtg ctcgggtcgg
     3061 ctcacgaggg ctgcctcccg ccgttcccag cggacgtttt catgattcct cagtacgggt
     3121 atctgacgct taatgatgga agccaggccg tgggtcgttc gtccttttac tgcctggaat
     3181 atttcccgtc gcaaatgcta agaacgggta acaacttcca gttcagctac gagtttgaga
     3241 acgtaccttt ccatagcagc tacgctcaca gccaaagcct ggaccgacta atgaatccac
     3301 tcatcgacca atacttgtac tatctctcta gaactattac cggttgggtt caaaaccaag
     3361 gaatacttcc gggtatggtt tggcaggaca gagatgtgta cctgcaagga cccatttggg
     3421 ccaaaattcc tcacacggac ggcaactttc acccttctcc gctgatggga gggtttggaa
     3481 tgaagcaccc gcctcctcag atcctcatca aaaacacacc tgtacctgcg gatcctccaa
     3541 cggccttcaa caaggacaag ctgaactctt tcatcaccca gtattctact ggccaagtca
     3601 gcgtggagat cgagtgggag ctgcagaagg aaaacagcaa gcgctggaac ccggagatcc
     3661 agtacacttc caactattac aagtctaata atgttgaatt tgctgttaat actgaaggtg
     3721 tatatagtga accccgcccc attggcacca gatacctgac tcgtaatctg taagtcgact
     3781 accgttcgta tagcatacat tatacgaagt tatcatatgt tcgagcagac atgataagat
     3841 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3901 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3961 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4021 gcaagtaaaa cctctacaaa tgtggtaaaa tcgagctcta ccgttcgtat aatgtatgct
     4081 atacgaagtt atgatatcaa gcttaggaac ccctagtgat ggagttggcc actccctctc
     4141 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4201 cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaag ctagcgggcg
     4261 attaaggaaa gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt
     4321 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4381 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4441 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4501 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4561 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4621 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4681 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     4741 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4801 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4861 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4921 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4981 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5041 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5101 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5161 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5221 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5281 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5341 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5401 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5461 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5521 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5581 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5641 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5701 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5761 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5821 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5881 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5941 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6001 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6061 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt aataaacaca
     6181 cacacaccaa caaccgtggt tggttgttgt gttggtttat tctcgag
//
