LOCUS       sequence-364480-        8543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-standard AAV2 rep-AAV-MaCPNS1 cap plasmid with AAV cap 
            expression controlled by a tTA-TRE amplification system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8543)
  AUTHORS   Chen X, Ravindra Kumar S, Adams CD, Yang D, Wang T, Wolfe DA, 
            Arokiaraj CM, Ngo V, Campos LJ, Griffiths JA, Ichiki T, Mazmanian 
            SK, Osborne PB, Keast JR, Miller CT, Fox AS, Chiu IM, Gradinaru V
  TITLE     Engineered AAVs for non-invasive gene delivery to rodent and 
            non-human primate nervous systems.
  JOURNAL   Neuron. 2022 Jul 20;110(14):2242-2257.e6. doi: 
            10.1016/j.neuron.2022.05.003. Epub 2022 May 27.
  PUBMED    35643078
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: 
            "10.1016/j.neuron.2022.05.003"; journalName: "Neuron"; date: 
            "2022-07-20- 20"; volume: "110"; issue: "14"; pages: "2242-2257.e6"
COMMENT     Sequence Label: pUCmini-iCAP-AAV.MaCPNS1
FEATURES             Location/Qualifiers
     source          1..8543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     153..170
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     154..170
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     180..199
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        180..198
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2788..2898
                     /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(2937..2955)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2938..2955)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2975..2992)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2981..3205
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3267..3285
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3304..3322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3341..3359
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3378..3396
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3415..3433
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3452..3470
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3489..3507
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3526..3544
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             6261..6272
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     rep_origin      complement(6828..7416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6908..6927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7587..8378,8379..8447))
                     /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     8210..8229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8448..8543,1..9))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc ccaatgatac gcgtcggtgc
       61 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      121 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
      181 aatacgactc actatagggc gaattgggta catcgacggt atcgggggag ctcgcagggt
      241 ctccattttg aagcgggagg tttgaacgcg cagccgccat gccggggttt tacgagattg
      301 tgattaaggt ccccagcgac cttgacgagc atctgcccgg catttctgac agctttgtga
      361 actgggtggc cgagaaggaa tgggagttgc cgccagattc tgacatggat ctgaatctga
      421 ttgagcaggc acccctgacc gtggccgaga agctgcagcg cgactttctg acggaatggc
      481 gccgtgtgag taaggccccg gaggctcttt tctttgtgca atttgagaag ggagagagct
      541 acttccacat gcacgtgctc gtggaaacca ccggggtgaa atccatggtt ttgggacgtt
      601 tcctgagtca gattcgcgaa aaactgattc agagaattta ccgcgggatc gagccgactt
      661 tgccaaactg gttcgcggtc acaaagacca gaaatggcgc cggaggcggg aacaaggtgg
      721 tggatgagtg ctacatcccc aattacttgc tccccaaaac ccagcctgag ctccagtggg
      781 cgtggactaa tatggaacag tatttaagcg cctgtttgaa tctcacggag cgtaaacggt
      841 tggtggcgca gcatctgacg cacgtgtcgc agacgcagga gcagaacaaa gagaatcaga
      901 atcccaattc tgatgcgccg gtgatcagat caaaaacttc agccaggtac atggagctgg
      961 tcgggtggct cgtggacaag gggattacct cggagaagca gtggatccag gaggaccagg
     1021 cctcatacat ctccttcaat gcggcctcca actcgcggtc ccaaatcaag gctgccttgg
     1081 acaatgcggg aaagattatg agcctgacta aaaccgcccc cgactacctg gtgggccagc
     1141 agcccgtgga ggacatttcc agcaatcgga tttataaaat tttggaacta aacgggtacg
     1201 atccccaata tgcggcttcc gtctttctgg gatgggccac gaaaaagttc ggcaagagga
     1261 acaccatctg gctgtttggg cctgcaacta ccgggaagac caacatcgcg gaggccatag
     1321 cccacactgt gcccttctac gggtgcgtaa actggaccaa tgagaacttt cccttcaacg
     1381 actgtgtgga caagatggtg atctggtggg aggaggggaa gatgaccgcc aaggtcgtgg
     1441 agtcggccaa agccattctc ggaggaagca aggtgcgcgt ggaccagaaa tgcaagtcct
     1501 cggcccagat agacccgact cccgtgatcg tcacctccaa caccaatatg tgcgccgtga
     1561 ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt gcaagaccgg atgttcaaat
     1621 ttgaactcac ccgccgtctg gatcatgact ttgggaaggt caccaagcag gaagtcaaag
     1681 actttttccg gtgggcaaag gatcacgtgg ttgaggtgga gcatgaattc tacgtcaaaa
     1741 agggtggagc caagaaaaga cccgccccca gtgacgcaga tataagtgag cccaaacggg
     1801 tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga agcttcgatc aactacgcgg
     1861 acaggtacca aaacaaatgt tctcgtcacg tgggcatgaa tctgatgctg tttccctgca
     1921 gacaatgcga gagactgaat cagaattcaa atatctgctt cactcacggt gtcaaagact
     1981 gtttagagtg ctttcccgtg tcagaatctc aacccgtttc tgtcgtcaaa aaggcgtatc
     2041 agaaactgtg ctacattcat cacatcatgg gaaaggtgcc agacgcttgc actgcttgcg
     2101 acctggtcaa tgtggacttg gatgactgtg tttctgaaca ataaatgact taaaccaggt
     2161 atgagtcggc tggataaatc taaagtcata aacggcgctc tggaattact caatgaagtc
     2221 ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga gcagcctacc
     2281 ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctggccat cgagatgctg
     2341 gacaggcatc atacccactt ctgccccctg gaaggcgagt catggcaaga ctttctgcgg
     2401 aacaacgcca agtcattccg ctgtgctctc ctctcacatc gcgacggggc taaagtgcat
     2461 ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg
     2521 tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt gggccacttt
     2581 acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga aagagagaca
     2641 cctaccaccg attctatgcc cccacttctg agacaagcaa ttgagctgtt cgaccggcag
     2701 ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct ggagaaacag
     2761 ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg attttgactt agacatgctc
     2821 ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc tcttgacgat
     2881 tttgaccttg acatgctccc cgggtaaatg catgaattcg atctagaggg ccctattcta
     2941 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3001 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3061 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3121 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3181 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgaat
     3241 caagctatca agtgccacct gacgtctccc tatcagtgat agagaagtcg acacgtctcg
     3301 agctccctat cagtgataga gaaggtacgt ctagaacgtc tccctatcag tgatagagaa
     3361 gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt acgtctagaa cgtctcccta
     3421 tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtacgtct
     3481 agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata
     3541 gagaaggtac cccctatata agcagagaga tctgttcaaa tttgaactga ctaagcggct
     3601 cccgccagat tttggcaaga ttactaagca ggaagtcaag gacttttttg cttgggcaaa
     3661 ggtcaatcag gtgccggtga ctcacgagtt taaagttccc agggaattgg cgggaactaa
     3721 aggggcggag aaatctctaa aacgcccact gggtgacgtc accaatacta gctataaaag
     3781 tctggagaag cgggccaggc tctcatttgt tcccgagacg cctcgcagtt cagacgtgac
     3841 tgttgatccc gctcctctgc gaccgctagc ttcgatcaac tacgcagaca ggtaccaaaa
     3901 caagtgttct cgtcacgtgg gcattaatct gattctgttt ccctgcagac aatgcgagag
     3961 aatgaatcag aactcaaata tctgcttcac tcacggacag aaagactgtt tagagtgctt
     4021 tcccgtgtca gaatctcaac ccgtttctgt cgtcaaaaag gcgtatcaga aactgtgcta
     4081 cattcatcat atcatgggaa aggtgccaga cgcttgcact gcctgcgatc tggtcaatgt
     4141 ggatttggat gactgcatct ttgaacaata aatgacttaa gccaggtatg gctgccgatg
     4201 gttatcttcc agattggctc gaggacaacc ttagtgaagg aattcgcgag tggtgggctt
     4261 tgaaacctgg agcccctcaa cccaaggcaa atcaacaaca tcaagacaac gctagaggtc
     4321 ttgtgcttcc gggttacaaa taccttggac ccggcaacgg actcgacaag ggggagccgg
     4381 tcaacgcagc agacgcggcg gccctcgagc acgacaaagc ctacgaccag cagctcaagg
     4441 ccggagacaa cccgtacctc aagtacaacc acgccgacgc cgagttccag gagcggctca
     4501 aagaagatac gtcttttggg ggcaacctcg ggcgagcagt cttccaggcc aaaaagaggc
     4561 ttcttgaacc tcttggtctg gttgaggaag cggctaagac ggctcctgga aagaagaggc
     4621 ctgtagagca gtctcctcag gaaccggact cctccgcggg tattggcaaa tcgggtgcac
     4681 agcccgctaa aaagagactc aatttcggtc agactggcga cacagagtca gtcccagacc
     4741 ctcaaccaat cggagaacct cccgcagccc cctcaggtgt gggatctctt acaatggctt
     4801 caggtggtgg cgcaccagtg gcagacaata acgaaggtgc cgatggagtg ggtagttcct
     4861 cgggaaattg gcattgcgat tcccaatggc tgggggacag agtcatcacc accagcaccc
     4921 gaacctgggc cctgcccacc tacaacaatc acctctacaa gcaaatctcc aacagcacat
     4981 ctggaggatc ttcaaatgac aacgcctact tcggctacag caccccctgg gggtattttg
     5041 acttcaacag attccactgc cacttctcac cacgtgactg gcagcgactc atcaacaaca
     5101 actggggatt ccggcctaag cgactcaact tcaagctctt taacattcag gtcaaagagg
     5161 ttacggacaa caatggagtc aagaccatcg ccaataacct taccagcacg gtccaggtct
     5221 tcacggactc agactatcag ctcccgtacg tgctcgggtc ggctcacgag ggctgcctcc
     5281 cgccgttccc agcggacgtt ttcatgattc ctcagtacgg gtatctgacg cttaatgatg
     5341 gaagccaggc cgtgggtcgt tcgtcctttt actgcctgga atatttcccg tcgcaaatgc
     5401 taagaacggg taacaacttc cagttcagct acgagtttga gaacgtacct ttccatagca
     5461 gctacgctca cagccaaagc ctggaccgac taatgaatcc actcatcgac caatacttgt
     5521 actatctctc tagaactatt aacggttctg gacagaatca acaaacgcta aaattcagtg
     5581 tggccggacc cagcaacatg gctgtccagg gaagaaacta catacctgga cccagctacc
     5641 gacaacaacg tgtctcaacc actgtgactc aaaacaacaa cagcgaattt gcttggcctg
     5701 gagcttcttc ttgggctctc aatggacgta atagcttgat gaatcctgga cctgctatgg
     5761 cctctcacaa agaaggagag gaccgtttct ttcctttgtc tggatcttta atttttggca
     5821 aacaaggtac tggcagagac aacgtggatg cggacaaagt catgataacc aacgaagaag
     5881 aaattaaaac tactaacccg gtagcaacgg agtcctatgg acaagtggcc acaaaccacc
     5941 agagtgccca accccacgaa ggcagcagca gagcacaggc gcagaccggt tgggttcaaa
     6001 accaaggaat acttccgggt atggtttggc aggacagaga tgtgtacctg caaggaccca
     6061 tttgggccaa aattcctcac acggacggca actttcaccc ttctccgctg atgggagggt
     6121 ttggaatgaa gcacccgcct cctcagatcc tcatcaaaaa cacacctgta cctgcggatc
     6181 ctccaacggc cttcaacaag gacaagctga actctttcat cacccagtat tctactggtc
     6241 aagtcagcgt ggagatcgag tgggagctgc agaaggaaaa cagcaagcgc tggaacccgg
     6301 agatccagta cacttccaac tattacaagt ctaataatgt tgaatttgct gttaatactg
     6361 aaggtgtata tagtgaaccc cgccccattg gcaccagata cctgactcgt aatctgtaag
     6421 tcgacttgct tgttaatcaa taaaccgttt aattcgtttc agttgaactt tggtctctgc
     6481 gaagggcaat tcgtttaaac ctgcaggact agaggtcctg tattagaggt cacgtgagtg
     6541 ttttgcgaca ttttgcgaca ccatgtggtc acgctgggta tttaagcccg agtgagcacg
     6601 cagggtctcc attttgaagc gggaggtttg aacgcgcagc cgccaagccg aattctgcag
     6661 atatcacatg tcctaggaac tatcgatcca tcacactggc ggccgctcga ctagagcggc
     6721 cgccaccgcg gtggagctcc agcttttgcg gaccgaatcg gaaagaacat gtgagcaaaa
     6781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7021 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7261 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7381 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7441 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7501 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7561 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7621 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7681 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7741 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7801 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7861 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7921 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7981 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8041 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8101 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8161 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8221 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8281 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8341 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8401 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8461 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8521 atttagaaaa ataaacaaat agg
//
