LOCUS       sequence-139644-        7401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified plasmid with a different backbone that yields higher titers
            of AAV-AS vectors. Encodes VP2-AS chimeric protein with 19 alanines 
            fused to N-terminus of AAV9.47 VP2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  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 7401)
  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: pGG-9.47_VP2_AS #2562
FEATURES             Location/Qualifiers
     source          1..7401
                     /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"
     primer_bind     complement(4469..4489)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4469..4487)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4508..4524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4508..4524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4521..4543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4532..4548
                     /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(4556..4562,4563..4580,4581..4586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4601..4622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4739..4756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4910..5498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4990..5009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5669..6460,6461..6529))
                     /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     6292..6311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6530..6634)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6660..7115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6797..6818)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7009..7028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7242..7264
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7256..7273
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7257..7273
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7283..7302
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7283..7301
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7330..7346
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7331..7347
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga gcatctgccc
       61 ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt gccgccagat
      121 tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag
      181 cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggctct 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 ggacaggtac caaaacaaat gttctcgtca cgtgggcatg
     1621 aatctgatgc tgtttccctg cagacaatgc gagagactga atcagaattc aaatatctgc
     1681 ttcactcacg gtgtcaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt
     1741 tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcacatcat gggaaaggtg
     1801 ccagacgctt gcactgcttg cgacctggtc aatgtggact tggatgactg tgtttctgaa
     1861 caataaatga cttaaaccag 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 tgtaattgct tgttaatcaa taaaccgttt aattcgtttc
     4201 agttgaactt tggtctctgc gaagggcgaa ttcgtttaaa cctgcaggac tagagtcctg
     4261 tattagaggt cacgtgagtg ttttgcgaca ttttgcgaca ccatgtggtc acgctgggta
     4321 tttaagcccg agtgagcacg cagggtctcc attttgaagc gggaggtttg aacgcgcagc
     4381 cgccaagccg aattctgcag atatccatca cactggcggc cgctcgacta gagcggccgc
     4441 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     4501 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4561 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4621 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4681 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4741 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4801 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4861 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4921 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4981 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5041 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5101 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5161 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5221 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5281 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5341 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5401 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5461 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5521 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5581 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5641 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5701 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5761 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5821 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5881 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5941 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6001 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6061 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6121 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6181 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6241 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6301 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6361 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6421 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6481 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6541 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6601 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     6661 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     6721 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6781 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6841 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6901 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6961 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7021 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7081 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     7141 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     7201 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     7261 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
     7321 ccccctcgat cgaggtcgac ggtatcgggg gagctcgcag ggtctccatt ttgaagcggg
     7381 aggtttgaac gcgcagccgc c
//
