LOCUS       sequence-146457-        7469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trans plasmid for production of recombinant AAV-B1 vectors. Plasmid 
            encodes AAV2 Rep and AAV-B1 capsid.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther. 
            2016 Apr 27. doi: 10.1038/mt.2016.84."
COMMENT     Sequence Label: pAR-B1(1986.1)
FEATURES             Location/Qualifiers
     source          1..7469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(4261..4267,4268..4285,4286..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(join(5374..6165,6166..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
//
