LOCUS Exported 7469 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Trans plasmid for production of recombinant AAV-B1 vectors. Plasmid encodes AAV2 Rep and AAV-B1 capsid. ACCESSION . VERSION . KEYWORDS pAR-B1(1986.1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7469) AUTHORS Choudhury SR, Fitzpatrick Z, Harris AF, Maitland SA, Ferreira JS, Zhang Y, Ma S, Sharma RB, Gray-Edwards HL, Johnson JA, Johnson AK, Alonso LC, Punzo C, Wagner KR, Maguire CA, Kotin RM, Martin DR, Sena-Esteves M TITLE In vivo selection yields AAV-B1 capsid for CNS and muscle gene therapy. JOURNAL Mol Ther. 2016 Apr 27. doi: 10.1038/mt.2016.84. PUBMED 27117222 REFERENCE 2 (bases 1 to 7469) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7469 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 3938..3949 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" promoter complement(4174..4192) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4213..4229) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4213..4229) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4226..4248) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4237..4253 /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(4261..4291) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4306..4327 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4444..4461) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4615..5203) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4695..4714) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5374..6234) /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 5997..6016 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6235..6339) /gene="bla" /label=AmpR promoter primer_bind 6407..6425 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6463..6485) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6585..6604 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(6671..7126) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6808..6829) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7020..7039 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7256..7278 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7270..7287 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7271..7287 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 7294..7313 /label=T7 /note="T7 promoter, forward primer" promoter 7294..7312 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 acgccggggt tttacgagat tgtgattaag gtccccagcg accttgacgg 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 caacctctct gagggcattc gcgagtggtg ggacttgaaa cctggagccc cgaagcccaa 1981 agccaaccag caaaagcagg acgacggccg gggtctggtg cttcctggct acaagtacct 2041 cggacccttc aacggactcg acaaggggga gcccgtcaac gcggcggacg cagcggccct 2101 cgagcacgac aaggcctacg accagcagct caaagcgggt gacaatccgt acctgcggta 2161 taaccacgcc gacgtcgagt ttcaggagcg tctgcaagaa gatacgtctt ttgggggcaa 2221 cctcgggcga gcagtcttcc aggccaagaa gagggttctc gaaccttttg gtctggttga 2281 ggaaggcgct aagacggctc ctggaaagaa acgtccggta gagcagtcgc cacaagagcc 2341 agactcctcc tcgggcatcg gcaagaaagg ccaacagccc gccagaaaaa gactcaattt 2401 tggccagact ggcgactcan agtcagttcc agaccctcaa cctctcggag aacctccagc 2461 agcgccctct ggtgtgggac ctaatacaat ggctgcaggc ggtggcgcac caatggcaga 2521 caataacgag ggcgccgacg gagtgngtaa ttcctcggga aattggcatt gcgattccac 2581 atggatgggg gacagagtca tcaccaccag cacccgaacc tgggccctgc ccacctacaa 2641 caaccacctc tacaagcaaa tctccaacgg cacctcggga ggaagcacca acgacaacac 2701 ctattttggc tacagcaccc cctgggggta ttttgatttc aacagattcc actgccactt 2761 ctcaccacgt gactggcagc gactcatcaa caacaactgg ggattccggc caaaaagact 2821 cagcttcaag ctcttcaaca tccaggtcaa ggaggtcacg cagaatgaag gcaccaagac 2881 catcgccaat aaccttacca gcacgattca ggtatttacg gactcggaat accagctgcc 2941 atacgtcctc ggctccgcgc accagggctg cctgcctccg ttcccggcgg acgtcttcat 3001 gattccccag tacggctacc ttacactgaa caatggaagt caagccgtag gccgttcctc 3061 cttctactgc ctggaatatt ttccttctca aatgctgaga acgggcaaca actttgagtt 3121 cagctaccag tttgaggacg tgccttttca cagcagctac gcgcacagcc aaagcctgga 3181 ccggctgatg aaccccctca tcgaccagta cctgtactac ctgtctcgga ctcagtccac 3241 gggaggtacc gcaagaactc agcagttgct attttctcag gccgggccta atacaatggc 3301 caatcaggca aagaactggc tgccaggacc ctgttaccgc caacaacgcg tctcaacgac 3361 aaccgggcaa aacaacaata gcaactctgc ctggactgct gggaccaaat accatctgaa 3421 tggaagaaat tcattggcta atcctggcat cgctatggca acacacaaag acgacgagga 3481 gcgttttttt cccagtaacg ggatcctgat ttttggcaaa caaaatgctg ccagagacaa 3541 tgcggattac agcgatgtca tgctcaccag cgaggaagaa atcaaaacca ctaaccctgt 3601 ggctacagag gaatacggta tcgtggcaga taacttgcag cagcaaaaca cggctcctca 3661 aattggaact gtcaacagcc agggggcctt acccggtatg gtctggcaga accgggacgt 3721 gtacctgcag ggtcccatct gggccaagat tcctcacacg gacggcaact tccacccatc 3781 tccgctgatg ggcggctttg gcctgaaaca tcctccgcct cagatcctga tcaagaacac 3841 gcctgtacct gcggatcctc cgaccacctt caaccagtca aagctgaact ctttcatcac 3901 gcaatacagc accggacagg tcagcgtgga aattgaatgg gagctacaga aggaaaacag 3961 caagcgctgg aaccccgaga tccagtacac ctccaactac tacaaatcta caagtgtgga 4021 ctttgctgtt aatacagaag gcgtgtactc tgaaccccgc cccattggca cccgttacct 4081 cacccgtaat ctgtaattgc ctgttaatca ataaaccggt tgattcgttt cagttgaact 4141 ttggtgtcgc ggccgctcga taagcttttg ttccctttag tgagggttaa ttccgagctt 4201 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4261 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4321 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 4381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 4441 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 4501 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 4561 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 4621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 4681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 4741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 4801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 4861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 4921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 4981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5041 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 6361 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 6421 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 6481 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 6541 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 6601 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 6661 ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa 6721 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa 6781 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac 6841 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa 6901 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct 6961 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gaggaaggaa 7021 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc 7081 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg ccattcgcca 7141 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 7201 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 7261 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattc 7321 gagctcggta cccctagagg tcctgtatta gaggtcacgt gagtgttttg cgacattttg 7381 cgacaccatg tggtcacgct gggtatttaa gcccgagtga gcacgcaggg tctccatttt 7441 gaagcgggag gtttgaacgc gcagccacc //