LOCUS       Exported                7462 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Original plasmid described in Choudhury et al. 2015 Mol Ther. for 
            production of AAV-AS vectors. Encodes VP1 and VP3 of AAV9.47.
ACCESSION   .
VERSION     .
KEYWORDS    pAR-9.47_VP1,3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  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 7462)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7462
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             3935..3946
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(4171..4189)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4210..4226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4210..4226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4223..4245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4234..4250
                     /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(4258..4288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4303..4324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4441..4458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4612..5200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4692..4711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5371..6231)
                     /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     5994..6013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6232..6336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6404..6422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6460..6482)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6582..6601
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6668..7123)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6805..6826)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7017..7036
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7249..7271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7263..7280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7264..7280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7287..7306
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7287..7305
                     /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 gtatggctgc 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 aaggcggctc ctggaaagaa gaggcctgta gagcagtctc ctcaggaacc
     2341 ggactcctcc gcgggtattg gcaaatcggg tgcacagccc gctaaaaaga gactcaattt
     2401 cggtcagact ggcgacacag agtcagtccc agaccctcaa ccaatcggag aacctcccgc
     2461 agccccctca ggtgtgggat ctcttacaat ggcttcaggt ggtggcgcac cagtggcaga
     2521 caataacgaa ggtgccgatg gagtgggtag ttcctcggga aattggcatt gcgattccca
     2581 atggctgggg gacagagtca tcaccaccag cacccgaacc tgggccctgc ccacctacaa
     2641 caatcacctc tacaagcaaa tctccaacag cacatctgga ggatcttcaa atgacaacgc
     2701 ctacttcggc tacagcaccc cctgggggta ttttgacttc aacagattcc actgccactt
     2761 ctcaccacgt gactggcagc gactcatcaa caacaactgg ggattccggc ctaagcgact
     2821 caacttcaag ctcttcaaca ttcaggtcaa agaggttacg gacaacaatg gagtcaagac
     2881 catcgccaat aaccttacca gcacggtcca ggtcttcacg gactcagact atcagctccc
     2941 gtacgtgctc gggtcggctc acgagggctg cctcccgccg ttcccagcgg acgttttcat
     3001 gattcctcag tacgggtatc tgacgcttaa tgatggaagc caggccgtgg gtcgttcgtc
     3061 cttttactgc ctggaatatt tcccgtcgca aatgctaaga acgggtaaca acttccagtt
     3121 caactacgag tttgagaacg tacctttcca tagcagctac gctcacagcc aaagcctgga
     3181 ccgactaatg aatccactca tcgaccaata cttgtactat ctctcaaaga ctattaacga
     3241 ttctggacag aatcaacaaa cgctaaaatt cagtgtggcc ggacccagca acatggctgt
     3301 ccagggaaga aactacatac ctggacccag ctaccgacaa caacgtgtct caaccactgt
     3361 gactcaaaac aacaacagcg aatttgcttg gcctggagct tcttcttggg ctctcaatgg
     3421 acgtaatagc ttgatgaatc ctggacctgc tatggccagc cacaaagaag gagaggaccg
     3481 tttctttcct ttgtctggat ctttaatttt tggcaaacaa ggaactggaa gagacaacgt
     3541 ggatgcggac gaagtcatga taaccaacga agaagaaatt aaaactacta acccggtagc
     3601 aacggagtcc tatggacaag tggccataaa ccaccagagt gcccaagcac aggcgcagac
     3661 cggctgggtt caaaaccaag gaatacttcc gggtatggtt tggcaggaca gagatgtgta
     3721 cctgcaagga cccatttggg ccaaaattcc tcacacggac ggcaactttc acccttctcc
     3781 gctgatggga gggtttggaa tgaagcaccc gcctcctcag atcctcatca aaaacacacc
     3841 tgtacctgcg gatcctccaa cggccttcaa caaggacaag ctgaactctt tcatcaccca
     3901 gtattctact ggccaagtca gcgtggagat cgagtgggag ctgcagaagg aaaacagcaa
     3961 gcgctggaac ccggagatcc agtacacttc caactattac aagtctaata atgttgaatt
     4021 tgctgttaat actgaaggtg tatatagtga accccgcccc attggcacca gatacctgac
     4081 tcgtaatctg taattacgtg ttaatcaata aaccggttga ttcgtttcag ttgaactttg
     4141 gtgtcgcggc cgctcgataa gcttttgttc cctttagtga gggttaattt cgagcttggc
     4201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6361 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6421 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6481 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6541 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6601 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6661 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6721 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6781 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6841 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6901 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6961 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     7021 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     7081 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     7141 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     7201 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     7261 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttcgagctcg
     7321 gtacccctag aggtcctgta ttagaggtca cgtgagtgtt ttgcgacatt ttgcgacacc
     7381 atgtggtcac gctgggtatt taagcccgag tgagcacgca gggtctccat tttgaagcgg
     7441 gaggtttgaa cgcgcagcca cc
//
