LOCUS       sequence-347131-        8543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-standard AAV2 rep-AAV.CAP-B10 cap plasmid with AAV cap 
            expression controlled by a tTA-TRE amplifcation system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8543)
  AUTHORS   Goertsen D, Flytzanis NC, Goeden N, Chuapoco MR, Cummins A, Chen Y, 
            Fan Y, Zhang Q, Sharma J, Duan Y, Wang L, Feng G, Chen Y, Ip NY, 
            Pickel J, Gradinaru V
  TITLE     AAV capsid variants with brain-wide transgene expression and 
            decreased liver targeting after intravenous delivery in mouse and 
            marmoset.
  JOURNAL   Nat Neurosci. 2022 Jan;25(1):106-115. doi: 
            10.1038/s41593-021-00969-4. Epub 2021 Dec 9.
  PUBMED    34887588
REFERENCE   2  (bases 1 to 8543)
  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"; doi: 
            "10.1038/s41593-021-00969-4"; journalName: "Nat Neurosci"; date: 
            "2022-01"; volume: "25"; issue: "1"; pages: "106-115"
COMMENT     Sequence Label: pUCmini-iCAP-AAV.CAP-B10
FEATURES             Location/Qualifiers
     source          1..8543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     130..152
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     144..161
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     145..161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     171..190
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        171..189
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2779..2889
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     promoter        complement(2928..2946)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2929..2946)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2966..2983)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2972..3196
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3258..3276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3295..3313
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3332..3350
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3369..3387
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3406..3424
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3443..3461
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3480..3498
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3517..3535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             6252..6263
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     rep_origin      complement(6819..7407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6899..6918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7578..8369,8370..8438))
                     /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     8201..8220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8439..8543)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cccaatgata cgcgtcggtg cgggcctctt
       61 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      121 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
      181 cactataggg cgaattgggt acatcgacgg tatcggggga gctcgcaggg tctccatttt
      241 gaagcgggag gtttgaacgc gcagccgcca tgccggggtt ttacgagatt gtgattaagg
      301 tccccagcga ccttgacgag catctgcccg gcatttctga cagctttgtg aactgggtgg
      361 ccgagaagga atgggagttg ccgccagatt ctgacatgga tctgaatctg attgagcagg
      421 cacccctgac cgtggccgag aagctgcagc gcgactttct gacggaatgg cgccgtgtga
      481 gtaaggcccc ggaggctctt ttctttgtgc aatttgagaa gggagagagc tacttccaca
      541 tgcacgtgct cgtggaaacc accggggtga aatccatggt tttgggacgt ttcctgagtc
      601 agattcgcga aaaactgatt cagagaattt accgcgggat cgagccgact ttgccaaact
      661 ggttcgcggt cacaaagacc agaaatggcg ccggaggcgg gaacaaggtg gtggatgagt
      721 gctacatccc caattacttg ctccccaaaa cccagcctga gctccagtgg gcgtggacta
      781 atatggaaca gtatttaagc gcctgtttga atctcacgga gcgtaaacgg ttggtggcgc
      841 agcatctgac gcacgtgtcg cagacgcagg agcagaacaa agagaatcag aatcccaatt
      901 ctgatgcgcc ggtgatcaga tcaaaaactt cagccaggta catggagctg gtcgggtggc
      961 tcgtggacaa ggggattacc tcggagaagc agtggatcca ggaggaccag gcctcataca
     1021 tctccttcaa tgcggcctcc aactcgcggt cccaaatcaa ggctgccttg gacaatgcgg
     1081 gaaagattat gagcctgact aaaaccgccc ccgactacct ggtgggccag cagcccgtgg
     1141 aggacatttc cagcaatcgg atttataaaa ttttggaact aaacgggtac gatccccaat
     1201 atgcggcttc cgtctttctg ggatgggcca cgaaaaagtt cggcaagagg aacaccatct
     1261 ggctgtttgg gcctgcaact accgggaaga ccaacatcgc ggaggccata gcccacactg
     1321 tgcccttcta cgggtgcgta aactggacca atgagaactt tcccttcaac gactgtgtgg
     1381 acaagatggt gatctggtgg gaggagggga agatgaccgc caaggtcgtg gagtcggcca
     1441 aagccattct cggaggaagc aaggtgcgcg tggaccagaa atgcaagtcc tcggcccaga
     1501 tagacccgac tcccgtgatc gtcacctcca acaccaatat gtgcgccgtg attgacggga
     1561 actcaacgac cttcgaacac cagcagccgt tgcaagaccg gatgttcaaa tttgaactca
     1621 cccgccgtct ggatcatgac tttgggaagg tcaccaagca ggaagtcaaa gactttttcc
     1681 ggtgggcaaa ggatcacgtg gttgaggtgg agcatgaatt ctacgtcaaa aagggtggag
     1741 ccaagaaaag acccgccccc agtgacgcag atataagtga gcccaaacgg gtgcgcgagt
     1801 cagttgcgca gccatcgacg tcagacgcgg aagcttcgat caactacgcg gacaggtacc
     1861 aaaacaaatg ttctcgtcac gtgggcatga atctgatgct gtttccctgt agacaatgcg
     1921 agagactgaa tcagaattca aatatctgct tcactcacgg tgtcaaagac tgtttagagt
     1981 gctttcccgt gtcagaatct caacccgttt ctgtcgtcaa aaaggcgtat cagaaactgt
     2041 gctacattca tcacatcatg ggaaaggtgc cagacgcttg cactgcttgc gacctggtca
     2101 atgtggactt ggatgactgt gtttctgaac aataaatgac ttaaaccagg tatgagtcgg
     2161 ctggataaat ctaaagtcat aaacggcgct ctggaattac tcaatgaagt cggtatcgaa
     2221 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg
     2281 cacgtgaaga acaagcgggc cctgctcgat gccctggcca tcgagatgct ggacaggcat
     2341 catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc
     2401 aagtcattcc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc
     2461 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa
     2521 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc
     2581 tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc
     2641 gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa
     2701 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc
     2761 gaaagcggcg ggccggccga cgcccttgac gattttgact tagacatgct cccagccgat
     2821 gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt
     2881 gacatgctcc ccgggtaaat gcatgaattc gatctagagg gccctattct atagtgtcac
     2941 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3001 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3061 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3121 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3181 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgaa tcaagctatc
     3241 aagtgccacc tgacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta
     3301 tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga agtcgacacg
     3361 tctcgagctc cctatcagtg atagagaagg tacgtctaga acgtctccct atcagtgata
     3421 gagaagtcga cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct
     3481 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     3541 ccccctatat aagcagagag atctgttcaa atttgaactg actaagcggc tcccgccaga
     3601 ttttggcaag attactaagc aggaagtcaa ggactttttt gcttgggcaa aggtcaatca
     3661 ggtgccggtg actcacgagt ttaaagttcc cagggaattg gcgggaacta aaggggcgga
     3721 gaaatctcta aaacgcccac tgggtgacgt caccaatact agctataaaa gtctggagaa
     3781 gcgggccagg ctctcatttg ttcccgagac gcctcgcagt tcagacgtga ctgttgatcc
     3841 cgctcctctg cgaccgctag cttcgatcaa ctacgcagac aggtaccaaa acaagtgttc
     3901 tcgtcacgtg ggcattaatc tgattctgtt tccctgcaga caatgcgaga gaatgaatca
     3961 gaactcaaat atctgcttca ctcacggaca gaaagactgt ttagagtgct ttcccgtgtc
     4021 agaatctcaa cccgtttctg tcgtcaaaaa ggcgtatcag aaactgtgct acattcatca
     4081 tatcatggga aaggtgccag acgcttgcac tgcctgcgat ctggtcaatg tggatttgga
     4141 tgactgcatc tttgaacaat aaatgactta agccaggtat ggctgccgat ggttatcttc
     4201 cagattggct cgaggacaac cttagtgaag gaattcgcga gtggtgggct ttgaaacctg
     4261 gagcccctca acccaaggca aatcaacaac atcaagacaa cgctagaggt cttgtgcttc
     4321 cgggttacaa ataccttgga cccggcaacg gactcgacaa gggggagccg gtcaacgcag
     4381 cagacgcggc ggccctcgag cacgacaaag cctacgacca gcagctcaag gccggagaca
     4441 acccgtacct caagtacaac cacgccgacg ccgagttcca ggagcggctc aaagaagata
     4501 cgtcttttgg gggcaacctc gggcgagcag tcttccaggc caaaaagagg cttcttgaac
     4561 ctcttggtct ggttgaggaa gcggctaaga cggctcctgg aaagaagagg cctgtagagc
     4621 agtctcctca ggaaccggac tcctccgcgg gtattggcaa atcgggtgca cagcccgcta
     4681 aaaagagact caatttcggt cagactggcg acacagagtc agtcccagac cctcaaccaa
     4741 tcggagaacc tcccgcagcc ccctcaggtg tgggatctct tacaatggct tcaggtggtg
     4801 gcgcaccagt ggcagacaat aacgaaggtg ccgatggagt gggtagttcc tcgggaaatt
     4861 ggcattgcga ttcccaatgg ctgggggaca gagtcatcac caccagcacc cgaacctggg
     4921 ccctgcccac ctacaacaat cacctctaca agcaaatctc caacagcaca tctggaggat
     4981 cttcaaatga caacgcctac ttcggctaca gcaccccctg ggggtatttt gacttcaaca
     5041 gattccactg ccacttctca ccacgtgact ggcagcgact catcaacaac aactggggat
     5101 tccggcctaa gcgactcaac ttcaagctct ttaacattca ggtcaaagag gttacggaca
     5161 acaatggagt caagaccatc gccaataacc ttaccagcac ggtccaggtc ttcacggact
     5221 cagactatca gctcccgtac gtgctcgggt cggctcacga gggctgcctc ccgccgttcc
     5281 cagcggacgt tttcatgatt cctcagtacg ggtatctgac gcttaatgat ggaagccagg
     5341 ccgtgggtcg ttcgtccttt tactgcctgg aatatttccc gtcgcaaatg ctaagaacgg
     5401 gtaacaactt ccagttcagc tacgagtttg agaacgtacc tttccatagc agctacgctc
     5461 acagccaaag cctggaccga ctaatgaatc cactcatcga ccaatacttg tactatctct
     5521 ctagaactat tgacggcgcc gccaccaaaa accaaacgct aaaattcagt gtggccggac
     5581 ccagcaacat ggctgtccag ggaagaaact acatacctgg acccagctac cgacaacaac
     5641 gtgtctcaac cactgtgact caaaacaaca acagcgaatt tgcttggcct ggagcttctt
     5701 cttgggctct caatggacgt aatagcttga tgaatcctgg acctgctatg gcctctcaca
     5761 aagaaggaga ggaccgtttc tttcctttgt ctggatcttt aatttttggc aaacaaggta
     5821 ctggcagaga caacgtggat gcggacaaag tcatgataac caacgaagaa gaaattaaaa
     5881 ctactaaccc ggtagcaacg gagtcctatg gacaagtggc cacaaaccac cagagtgatg
     5941 ggactttggc ggtgcctttt aaggcacagg cgcagaccgg ttgggttcaa aaccaaggaa
     6001 tacttccggg tatggtttgg caggacagag atgtgtacct gcaaggaccc atttgggcca
     6061 aaattcctca cacggacggc aactttcacc cttctccgct gatgggaggg tttggaatga
     6121 agcacccgcc tcctcagatc ctcatcaaaa acacacctgt acctgcggat cctccaacgg
     6181 ccttcaacaa ggacaagctg aactctttca tcacccagta ttctactggt caagtcagcg
     6241 tggagatcga gtgggagctg cagaaggaaa acagcaagcg ctggaacccg gagatccagt
     6301 acacttccaa ctattacaag tctaataatg ttgaatttgc tgttaatact gaaggtgtat
     6361 atagtgaacc ccgccccatt ggcaccagat acctgactcg taatctgtaa gtcgacttgc
     6421 ttgttaatca ataaaccgtt taattcgttt cagttgaact ttggtctctg cgaagggcaa
     6481 ttcgtttaaa cctgcaggac tagaggtcct gtattagagg tcacgtgagt gttttgcgac
     6541 attttgcgac accatgtggt cacgctgggt atttaagccc gagtgagcac gcagggtctc
     6601 cattttgaag cgggaggttt gaacgcgcag ccgccaagcc gaattctgca gatatcacat
     6661 gtcctaggaa ctatcgatcc atcacactgg cggccgctcg actagagcgg ccgccaccgc
     6721 ggtggagctc cagcttttgc ggaccgaatc ggaaagaaca tgtgagcaaa aggccagcaa
     6781 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6841 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6901 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6961 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7021 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7081 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7141 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7201 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7261 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7321 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7381 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7441 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7501 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8521 aataaacaaa taggggttcc gcg
//
