LOCUS       sequence-329744-        7483 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV rep/cap plasmid. Capsid is mouse brain tropic AAV-F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  AUTHORS   Hanlon KS, Meltzer JC, Buzhdygan T, Cheng MJ, Sena-Esteves M, 
            Bennett RE, Sullivan TP, Razmpour R, Gong Y, Ng C, Nammour J, Maiz 
            D, Dujardin S, Ramirez SH, Hudry E, Maguire CA
  TITLE     Selection of an Efficient AAV Vector for Robust CNS Transgene 
            Expression.
  JOURNAL   Mol Ther Methods Clin Dev. 2019 Oct 23;15:320-332. doi: 
            10.1016/j.omtm.2019.10.007. eCollection 2019 Dec 13.
  PUBMED    31788496
REFERENCE   2  (bases 1 to 7483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Methods Clin Dev."; date: "2019-10-23"; volume: "15:320-332. doi"; 
            pages: " 10.1016/j.omtm.2019.10.007. eCollection 2019 Dec 13"
COMMENT     Sequence Label: AAV-F in pAR-9
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3957..3968
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(4193..4211)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4232..4248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4232..4248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4245..4267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4256..4272
                     /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(4280..4286,4287..4304,4305..4310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4325..4346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4463..4480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4634..5222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4714..4733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5393..6184,6185..6253))
                     /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     6016..6035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6254..6358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6426..6444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6482..6504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6604..6623
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6690..7145)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6827..6848)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7039..7058
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7271..7293
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7285..7302
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7286..7302
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7309..7328
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7309..7327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cacgccgggg ttttacgaga ttgtgattaa ggtccccagc gaccttgacg agcatctgcc
       61 cggcatttct gacagctttg tgaactgggt ggccgagaag gaatgggagt tgccgccaga
      121 ttctgacatg gatctgaatc tgattgagca ggcacccctg accgtggccg agaagctgca
      181 gcgcgacttt ctgacggaat ggcgccgtgt gagtaaggcc ccggaggccc ttttctttgt
      241 gcaatttgag aagggagaga gctacttcca catgcacgtg ctcgtggaaa ccaccggggt
      301 gaaatccatg gttttgggac gtttcctgag tcagattcgc gaaaaactga ttcagagaat
      361 ttaccgcggg atcgagccga ctttgccaaa ctggttcgcg gtcacaaaga ccagaaatgg
      421 cgccggaggc gggaacaagg tggtggatga gtgctacatc cccaattact tgctccccaa
      481 aacccagcct gagctccagt gggcgtggac taatatggaa cagtatttaa gcgcctgttt
      541 gaatctcacg gagcgtaaac ggttggtggc gcagcatctg acgcacgtgt cgcagacgca
      601 ggagcagaac aaagagaatc agaatcccaa ttctgatgcg ccggtgatca gatcaaaaac
      661 ttcagccagg tacatggagc tggtcgggtg gctcgtggac aaggggatta cctcggagaa
      721 gcagtggatc caggaggacc aggcctcata catctccttc aatgcggcct ccaactcgcg
      781 gtcccaaatc aaggctgcct tggacaatgc gggaaagatt atgagcctga ctaaaaccgc
      841 ccccgactac ctggtgggcc agcagcccgt ggaggacatt tccagcaatc ggatttataa
      901 aattttggaa ctaaacgggt acgatcccca atatgcggct tccgtctttc tgggatgggc
      961 cacgaaaaag ttcggcaaga ggaacaccat ctggctgttt gggcctgcaa ctaccgggaa
     1021 gaccaacatc gcggaggcca tagcccacac tgtgcccttc tacgggtgcg taaactggac
     1081 caatgagaac tttcccttca acgactgtgt cgacaagatg gtgatctggt gggaggaggg
     1141 gaagatgacc gccaaggtcg tggagtcggc caaagccatt ctcggaggaa gcaaggtgcg
     1201 cgtggaccag aaatgcaagt cctcggccca gatagacccg actcccgtga tcgtcacctc
     1261 caacaccaac atgtgcgccg tgattgacgg gaactcaacg accttcgaac accagcagcc
     1321 gttgcaagac cggatgttca aatttgaact cacccgccgt ctggatcatg actttgggaa
     1381 ggtcaccaag caggaagtca aagacttttt ccggtgggca aaggatcacg tggttgaggt
     1441 ggagcatgaa ttctacgtca aaaagggtgg agccaagaaa agacccgccc ccagtgacgc
     1501 agatataagt gagcccaaac gggtgcgcga gtcagttgcg cagccatcga cgtcagacgc
     1561 ggaagcttcg atcaactacg cagacaggta ccaaaacaaa tgttctcgtc acgtgggcat
     1621 gaatctgatg ctgtttccct gcagacaatg cgagagaatg aatcagaatt caaatatctg
     1681 cttcactcac ggacagaaag actgtttaga gtgctttccc gtgtcagaat ctcaacccgt
     1741 ttctgtcgtc aaaaaggcgt atcagaaact gtgctacatt catcatatca tgggaaaggt
     1801 gccagacgct tgcactgcct gcgatctggt caatgtggat ttggatgact gcatctttga
     1861 acaataaatg atttaaatca ggtatggctg ccgatggtta tcttccagat tggctcgagg
     1921 acaaccttag tgaaggaatt cgcgagtggt gggctttgaa acctggagcc cctcaaccca
     1981 aggcaaatca acaacatcaa gacaacgctc gaggtcttgt gcttccgggt tacaaatacc
     2041 ttggacccgg caacggactc gacaaggggg agccggtcaa cgcagcagac gcggcggccc
     2101 tcgagcacga caaggcctac gaccagcagc tcaaggccgg agacaacccg tacctcaagt
     2161 acaaccacgc cgacgccgag ttccaggagc ggctcaaaga agatacgtct tttgggggca
     2221 acctcgggcg agcagtcttc caggccaaaa agaggcttct tgaacctctt ggtctggttg
     2281 aggaagcggc taagacggct cctggaaaga agaggcctgt agagcagtct cctcaggaac
     2341 cggactcctc cgcgggtatt ggcaaatcgg gtgcacagcc cgctaaaaag agactcaatt
     2401 tcggtcagac tggcgacaca gagtcagtcc cagaccctca accaatcgga gaacctcccg
     2461 cagccccctc aggtgtggga tctcttacaa tggcttcagg tggtggcgca ccagtggcag
     2521 acaataacga aggtgccgat ggagtgggta gttcctcggg aaattggcat tgcgattccc
     2581 aatggctggg ggacagagtc atcaccacca gcacccgaac ctgggccctg cccacctaca
     2641 acaatcacct ctacaagcaa atctccaaca gcacatctgg aggatcttca aatgacaacg
     2701 cctacttcgg ctacagcacc ccctgggggt attttgactt caacagattc cactgccact
     2761 tctcaccacg tgactggcag cgactcatca acaacaactg gggattccgg cctaagcgac
     2821 tcaacttcaa gctcttcaac attcaggtca aagaggttac ggacaacaat ggagtcaaga
     2881 ccatcgccaa taaccttacc agcacggtcc aggtcttcac ggactcagac tatcagctcc
     2941 cgtacgtgct cgggtcggct cacgagggct gcctcccgcc gttcccagcg gacgttttca
     3001 tgattcctca gtacgggtat ctgacgctta atgatggaag ccaggccgtg ggtcgttcgt
     3061 ccttttactg cctggaatat ttcccgtcgc aaatgctaag aacgggtaac aacttccagt
     3121 tcagctacga gtttgagaac gtacctttcc atagcagcta cgctcacagc caaagcctgg
     3181 accgactaat gaatccactc atcgaccaat acttgtacta tctctcaaag actattaacg
     3241 gttctggaca gaatcaacaa acgctaaaat tcagtgtggc cggacccagc aacatggctg
     3301 tccagggaag aaactacata cctggaccca gctaccgaca acaacgtgtc tcaaccactg
     3361 tgactcaaaa caacaacagc gaatttgctt ggcctggagc ttcttcttgg gctctcaatg
     3421 gacgtaatag cttgatgaat cctggacctg ctatggccag ccacaaagaa ggagaggacc
     3481 gtttctttcc tttgtctgga tctttaattt ttggcaaaca aggaactgga agagacaacg
     3541 tggatgcgga caaagtcatg ataaccaacg aagaagaaat taaaactact aacccggtag
     3601 caacggagtc ctatggacaa gtggccacaa accaccagag tgcccaattt gttgttggtc
     3661 agagttatgc acaggcgcag accggctggg ttcaaaacca aggaatactt ccgggtatgg
     3721 tttggcagga cagagatgtg tacctgcaag gacccatttg ggccaaaatt cctcacacgg
     3781 acggcaactt tcacccttct ccgctgatgg gagggtttgg aatgaagcac ccgcctcctc
     3841 agatcctcat caaaaacaca cctgtacctg cggatcctcc aacggccttc aacaaggaca
     3901 agctgaactc tttcatcacc cagtattcta ctggccaagt cagcgtggag atcgagtggg
     3961 agctgcagaa ggaaaacagc aagcgctgga acccggagat ccagtacact tccaactatt
     4021 acaagtctaa taatgttgaa tttgctgtta atactgaagg tgtatatagt gaaccccgcc
     4081 ccattggcac cagatacctg actcgtaatc tgtaattacg tgttaatcaa taaaccggtt
     4141 gattcgtttc agttgaactt tggtgtcgcg gccgctcgat aagcttttgt tccctttagt
     4201 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4261 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4321 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4381 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4441 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5341 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5401 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5461 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5521 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5581 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5641 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5701 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5761 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5821 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5881 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5941 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6001 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6061 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6121 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6181 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6241 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6301 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6361 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6421 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6481 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6541 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6601 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     6661 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     6721 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6781 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6841 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6901 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6961 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     7021 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     7081 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7141 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7201 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7261 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     7321 ctatagggcg aattcgagct cggtacccct agaggtcctg tattagaggt cacgtgagtg
     7381 ttttgcgaca ttttgcgaca ccatgtggtc acgctgggta tttaagcccg agtgagcacg
     7441 cagggtctcc attttgaagc gggaggtttg aacgcgcagc cac
//
