LOCUS       sequence-404982-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rep and AAP protein expression for AAV CREATE libraries.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  AUTHORS   Chuapoco MR, Flytzanis NC, Goeden N, Christopher Octeau J, Roxas KM,
            Chan KY, Scherrer J, Winchester J, Blackburn RJ, Campos LJ, Man KNM,
            Sun J, Chen X, Lefevre A, Singh VP, Arokiaraj CM, Shay TF, 
            Vendemiatti J, Jang MJ, Mich JK, Bishaw Y, Gore BB, Omstead V, 
            Taskin N, Weed N, Levi BP, Ting JT, Miller CT, Deverman BE, Pickel 
            J, Tian L, Fox AS, Gradinaru V
  TITLE     Adeno-associated viral vectors for functional intravenous gene 
            transfer throughout the non-human primate brain.
  JOURNAL   Nat Nanotechnol. 2023 Jul 10. doi: 10.1038/s41565-023-01419-x.
  PUBMED    37430038
REFERENCE   2  (bases 1 to 6890)
  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: "Nat 
            Nanotechnol. 2023 Jul 10. doi: 10.1038/s41565-023-01419-x."
COMMENT     Sequence Label: AAV2/9 REP-AAP-Delta-CAP
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..73)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(733..1524,1525..1593))
                     /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     1356..1375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1594..1698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1724..2179)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1861..1882)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2073..2092
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2306..2328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2320..2337
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2321..2337
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2347..2366
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2347..2365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2394..2410
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2395..2411
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5960..5971
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(6423..6443)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6423..6441)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6462..6478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6462..6478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6475..6497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6486..6502
                     /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(6510..6516,6517..6534,6535..6540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6555..6576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6693..6710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6864..6890,1..562))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
       61 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
      841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
      901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
      961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1021 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1081 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1141 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1201 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1261 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1621 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1681 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     1741 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1801 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     1861 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1921 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1981 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2041 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2101 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2161 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2221 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2281 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     2341 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgatcgaggt
     2401 cgacggtatc gggggagctc gcagggtctc cattttgaag cgggaggttt gaacgcgcag
     2461 ccgccatgcc ggggttttac gagattgtga ttaaggtccc cagcgacctt gacgagcatc
     2521 tgcccggcat ttctgacagc tttgtgaact gggtggccga gaaggaatgg gagttgccgc
     2581 cagattctga catggatctg aatctgattg agcaggcacc cctgaccgtg gccgagaagc
     2641 tgcagcgcga ctttctgacg gaatggcgcc gtgtgagtaa ggccccggag gctcttttct
     2701 ttgtgcaatt tgagaaggga gagagctact tccacatgca cgtgctcgtg gaaaccaccg
     2761 gggtgaaatc catggttttg ggacgtttcc tgagtcagat tcgcgaaaaa ctgattcaga
     2821 gaatttaccg cgggatcgag ccgactttgc caaactggtt cgcggtcaca aagaccagaa
     2881 atggcgccgg aggcgggaac aaggtggtgg atgagtgcta catccccaat tacttgctcc
     2941 ccaaaaccca gcctgagctc cagtgggcgt ggactaatat ggaacagtat ttaagcgcct
     3001 gtttgaatct cacggagcgt aaacggttgg tggcgcagca tctgacgcac gtgtcgcaga
     3061 cgcaggagca gaacaaagag aatcagaatc ccaattctga tgcgccggtg atcagatcaa
     3121 aaacttcagc caggtacatg gagctggtcg ggtggctcgt ggacaagggg attacctcgg
     3181 agaagcagtg gatccaggag gaccaggcct catacatctc cttcaatgcg gcctccaact
     3241 cgcggtccca aatcaaggct gccttggaca atgcgggaaa gattatgagc ctgactaaaa
     3301 ccgcccccga ctacctggtg ggccagcagc ccgtggagga catttccagc aatcggattt
     3361 ataaaatttt ggaactaaac gggtacgatc cccaatatgc ggcttccgtc tttctgggat
     3421 gggccacgaa aaagttcggc aagaggaaca ccatctggct gtttgggcct gcaactaccg
     3481 ggaagaccaa catcgcggag gccatagccc acactgtgcc cttctacggg tgcgtaaact
     3541 ggaccaatga gaactttccc ttcaacgact gtgtcgacaa gatggtgatc tggtgggagg
     3601 aggggaagat gaccgccaag gtcgtggagt cggccaaagc cattctcgga ggaagcaagg
     3661 tgcgcgtgga ccagaaatgc aagtcctcgg cccagataga cccgactccc gtgatcgtca
     3721 cctccaacac caacatgtgc gccgtgattg acgggaactc aacgaccttc gaacaccagc
     3781 agccgttgca agaccggatg ttcaaatttg aactcacccg ccgtctggat catgactttg
     3841 ggaaggtcac caagcaggaa gtcaaagact ttttccggtg ggcaaaggat cacgtggttg
     3901 aggtggagca tgaattctac gtcaaaaagg gtggagccaa gaaaagaccc gcccccagtg
     3961 acgcagatat aagtgagccc aaacgggtgc gcgagtcagt tgcgcagcca tcgacgtcag
     4021 acgcggaagc ttcgatcaac tacgcggaca ggtaccaaaa caaatgttct cgtcacgtgg
     4081 gcatgaatct gatgctgttt ccctgcagac aatgcgagag actgaatcag aattcaaata
     4141 tctgcttcac tcacggtgtc aaagactgtt tagagtgctt tcccgtgtca gaatctcaac
     4201 ccgtttctgt cgtcaaaaag gcgtatcaga aactgtgcta cattcatcac atcatgggaa
     4261 aggtgccaga cgcttgcact gcttgcgacc tggtcaatgt ggacttggat gactgtgttt
     4321 ctgaacaata aatgacttaa accaggtatg gctgccgatg gttaacttcc agattgactc
     4381 gaggacaacc ttagtgaagg aattcgcgag tggtgggctt tgaaacctgg agcccctcaa
     4441 cccaaggcaa atcaacaaca tcaagacaac gctcgaggtc ttgtgcttcc gggttacaaa
     4501 taccttggac ccggcaacgg actcgacaag ggggagccgg tcaacgcagc agacgcggcg
     4561 gccctcgagc acgacaaggc ctacgaccag cagctcaagg ccggagacaa cccgtacctc
     4621 aagtacaacc acgccgacgc cgagttccag gagcggctca aagaagatac gtcttttggg
     4681 ggcaacctcg ggcgagcagt cttccaggcc aaaaagaggc ttcttgaacc tcttggtctg
     4741 gttgaggaag cggctaagac ggctcctgga tagaagaggc ctgtagagta gtctcctcag
     4801 gaaccggact cctccgcggg tattggcaaa tcgggtgcac agcccgctaa aaagagactc
     4861 aatttcggtc agactggcga cacagagtca gtcccagacc ctcaaccaat cggagaacct
     4921 cccgcagccc cctcaggtgt gggatctctt acaatggctt caggtggtgg cgcaccagtg
     4981 gcagacaata actaaggtgc cgatggagtg ggtagttcct cgggaaattg gcattgcgat
     5041 tcccaatggc tgggggacag agtcatcacc accagcaccc gaacctgggc cctgcccacc
     5101 tacaacaatc acctctacaa gcaaatctcc aacagcacat ctggaggatc ttcaaatgac
     5161 aacgcctact tcggctacag caccccctgg gggtattttg acttcaacag attccactgc
     5221 cacttctcac cacgtgactg gcagcgactc atcaacaaca actggggatt ccggcctaag
     5281 cgactcaact tcaagctctt caacattcag gtcaaagagg ttacggacaa caatggagtc
     5341 aagaccatcg ccaataacct taccagcacg gtccaggtct tcacggactc agactatcag
     5401 ctcccgtacg tgctcgggtc ggctcacgag ggctgcctcc cgccgttccc agcggacgtt
     5461 ttcatgattc ctcagtacgg gtatctgacg cttaatgatg gaagccaggc cgtgggtcgt
     5521 tcgtcctttt actgcctgga atatttcccg tcgcaaatgc taagaacggg taacaacttc
     5581 cagttcagct acgagtttga gaacgtacct ttccatagca gctacgctca cagccaaagc
     5641 ctggaccgac taatgaatcc actcatcgac caatacttgt actatctctt gggttcaaaa
     5701 ccaaggaata cttccgggta tggtttggca ggacagagat gtgtacctgc aaggacccat
     5761 ttgggccaaa attcctcaca cggacggcaa ctttcaccct tctccgctga tgggagggtt
     5821 tggaatgaag cacccgcctc ctcagatcct catcaaaaac acacctgtac ctgcggatcc
     5881 tccaacggcc ttcaacaagg acaagctgaa ctctttcatc acccagtatt ctactggcca
     5941 agtcagcgtg gagatcgagt gggagctgca gaaggaaaac agcaagcgct ggaacccgga
     6001 gatccagtac acttccaact attacaagtc taataatgtt gaatttgctg ttaatactga
     6061 aggtgtatat agtgaacccc gccccattgg caccagatac ctgactcgta atctgtaatt
     6121 gcttgttaat caataaaccg tttaattcgt ttcagttgaa ctttggtctc tgcgaagggc
     6181 gaattcgttt aaacctgcag gactagaggt cctgtattag aggtcacgtg agtgttttgc
     6241 gacattttgc gacaccatgt ggtcacgctg ggtatttaag cccgagtgag cacgcagggt
     6301 ctccattttg aagcgggagg tttgaacgcg cagccgccaa gccgaattct gcagatatcc
     6361 atcacactgg cggccgctcg actagagcgg ccgccaccgc ggtggagctc cagcttttgt
     6421 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6481 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6541 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6601 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6661 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
//
