LOCUS       sequence-141992-        7534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Original plasmid described in Choudhury et al, 2015 Mol. Ther. for 
            production of AAV-AS vectors. Encodes VP2_AS chimeric protein with 
            19 alanines fused to the N-terminus of AAV9.47 VP2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7534)
  AUTHORS   Choudhury SR, Harris AF, Cabral DJ, Keeler AM, Sapp E, Ferreira JS, 
            Gray-Edwards HL, Johnson JA, Johnson AK, Su Q, Stoica L, DiFiglia M,
            Aronin N, Martin DR, Gao G, Sena-Esteves M
  TITLE     Widespread Central Nervous System Gene Transfer and Silencing After 
            Systemic Delivery of Novel AAV-AS Vector.
  JOURNAL   Mol Ther. 2015 Dec 28. doi: 10.1038/mt.2015.231.
  PUBMED    26708003
REFERENCE   2  (bases 1 to 7534)
  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: "Mol Ther. 
            2015 Dec 28. doi: 10.1038/mt.2015.231."
COMMENT     Sequence Label: pAR-9.47_VP2_AS
FEATURES             Location/Qualifiers
     source          1..7534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4007..4018
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(4243..4261)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4282..4298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4282..4298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4295..4317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4306..4322
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4330..4336,4337..4354,4355..4360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4375..4396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4513..4530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4684..5272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4764..4783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5443..6234,6235..6303))
                     /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     6066..6085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6304..6408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6476..6494
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6532..6554)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6654..6673
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6740..7195)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6877..6898)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7089..7108
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7321..7343
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7335..7352
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7336..7352
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7359..7378
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7359..7377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga gcatctgccc
       61 ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt gccgccagat
      121 tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag
      181 cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct tttctttgtg
      241 caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac caccggggtg
      301 aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat tcagagaatt
      361 taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac cagaaatggc
      421 gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt gctccccaaa
      481 acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag cgcctgtttg
      541 aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc gcagacgcag
      601 gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag atcaaaaact
      661 tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac ctcggagaag
      721 cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc caactcgcgg
      781 tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac taaaaccgcc
      841 cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg gatttataaa
      901 attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct gggatgggcc
      961 acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac taccgggaag
     1021 accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt aaactggacc
     1081 aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg ggaggagggg
     1141 aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag caaggtgcgc
     1201 gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat cgtcacctcc
     1261 aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca ccagcagccg
     1321 ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga ctttgggaag
     1381 gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt ggttgaggtg
     1441 gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc cagtgacgca
     1501 gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac gtcagacgcg
     1561 gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca cgtgggcatg
     1621 aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc aaatatctgc
     1681 ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt
     1741 tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat gggaaaggtg
     1801 ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg catctttgaa
     1861 caataaatga tttaaatcag gtctggctgc cgatggttat cttccagatt ggctcgagga
     1921 caaccttagt gaaggaattc gcgagtggtg ggctttgaaa cctggagccc ctcaacccaa
     1981 ggcaaatcaa caacatcaag acaacgctcg aggtcttgtg cttccgggtt acaaatacct
     2041 tggacccggc aacggactcg acaaggggga gccggtcaac gcagcagacg cggcggccct
     2101 cgagcacgac aaggcctacg accagcagct caaggccgga gacaacccgt acctcaagta
     2161 caaccacgcc gacgccgagt tccaggagcg gctcaaagaa gatacgtctt ttgggggcaa
     2221 cctcgggcga gcagtcttcc aggccaaaaa gaggcttctt gaacctcttg gtctggttga
     2281 ggaagcggct aagatggcag ctgcggccgc cgcggctgca gcggctgcag ccgccgcagc
     2341 tgcggctgca gcgggcggcg gcggcagcgc tcctggaaag aagaggcctg tagagcagtc
     2401 tcctcaggaa ccggactcct ccgcgggtat tggcaaatcg ggtgcacagc ccgctaaaaa
     2461 gagactcaat ttcggtcaga ctggcgacac agagtcagtc ccagaccctc aaccaatcgg
     2521 agaacctccc gcagccccct caggtgtggg atctcttaca atggcttcag gtggtggcgc
     2581 accagtggca gacaataacg aaggtgccga tggagtgggt agttcctcgg gaaattggca
     2641 ttgcgattcc caatggctgg gggacagagt catcaccacc agcacccgaa cctgggccct
     2701 gcccacctac aacaatcacc tctacaagca aatctccaac agcacatctg gaggatcttc
     2761 aaatgacaac gcctacttcg gctacagcac cccctggggg tattttgact tcaacagatt
     2821 ccactgccac ttctcaccac gtgactggca gcgactcatc aacaacaact ggggattccg
     2881 gcctaagcga ctcaacttca agctcttcaa cattcaggtc aaagaggtta cggacaacaa
     2941 tggagtcaag accatcgcca ataaccttac cagcacggtc caggtcttca cggactcaga
     3001 ctatcagctc ccgtacgtgc tcgggtcggc tcacgagggc tgcctcccgc cgttcccagc
     3061 ggacgttttc atgattcctc agtacgggta tctgacgctt aatgatggaa gccaggccgt
     3121 gggtcgttcg tccttttact gcctggaata tttcccgtcg caaatgctaa gaacgggtaa
     3181 caacttccag ttcaactacg agtttgagaa cgtacctttc catagcagct acgctcacag
     3241 ccaaagcctg gaccgactaa tgaatccact catcgaccaa tacttgtact atctctcaaa
     3301 gactattaac gattctggac agaatcaaca aacgctaaaa ttcagtgtgg ccggacccag
     3361 caacatggct gtccagggaa gaaactacat acctggaccc agctaccgac aacaacgtgt
     3421 ctcaaccact gtgactcaaa acaacaacag cgaatttgct tggcctggag cttcttcttg
     3481 ggctctcaat ggacgtaata gcttgatgaa tcctggacct gctatggcca gccacaaaga
     3541 aggagaggac cgtttctttc ctttgtctgg atctttaatt tttggcaaac aaggaactgg
     3601 aagagacaac gtggatgcgg acgaagtcat gataaccaac gaagaagaaa ttaaaactac
     3661 taacccggta gcaacggagt cctatggaca agtggccata aaccaccaga gtgcccaagc
     3721 acaggcgcag accggctggg ttcaaaacca aggaatactt ccgggtatgg tttggcagga
     3781 cagagatgtg tacctgcaag gacccatttg ggccaaaatt cctcacacgg acggcaactt
     3841 tcacccttct ccgctgatgg gagggtttgg aatgaagcac ccgcctcctc agatcctcat
     3901 caaaaacaca cctgtacctg cggatcctcc aacggccttc aacaaggaca agctgaactc
     3961 tttcatcacc cagtattcta ctggccaagt cagcgtggag atcgagtggg agctgcagaa
     4021 ggaaaacagc aagcgctgga acccggagat ccagtacact tccaactatt acaagtctaa
     4081 taatgttgaa tttgctgtta atactgaagg tgtatatagt gaaccccgcc ccattggcac
     4141 cagatacctg actcgtaatc tgtaattacg tgttaatcaa taaaccggtt gattcgtttc
     4201 agttgaactt tggtgtcgcg gccgctcgat aagcttttgt tccctttagt gagggttaat
     4261 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4321 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4381 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4441 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4501 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4561 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4621 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4681 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4741 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4801 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4861 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4921 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4981 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5041 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5101 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5161 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5221 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6361 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6421 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6481 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6541 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6601 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6661 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6721 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6781 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6841 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6901 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6961 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     7021 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     7081 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     7141 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     7201 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7261 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7321 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     7381 aattcgagct cggtacccct agaggtcctg tattagaggt cacgtgagtg ttttgcgaca
     7441 ttttgcgaca ccatgtggtc acgctgggta tttaagcccg agtgagcacg cagggtctcc
     7501 attttgaagc gggaggtttg aacgcgcagc cacc
//
