LOCUS       sequence-259997-        8543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-standard AAV2 rep-AAV-PHP.B3 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   Deverman BE, Pravdo PL, Simpson BP, Kumar SR, Chan KY, Banerjee A, 
            Wu WL, Yang B, Huber N, Pasca SP, Gradinaru V
  TITLE     Cre-dependent selection yields AAV variants for widespread gene 
            transfer to the adult brain.
  JOURNAL   Nat Biotechnol. 2016 Feb;34(2):204-9. doi: 10.1038/nbt.3440. Epub 
            2016 Feb 1.
  PUBMED    26829320
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/nbt.3440"; 
            journalName: "Nat Biotechnol"; date: "2016-02"; volume: "34"; issue:
            "2"; pages: "204-9"
COMMENT     Sequence Label: pUCmini-iCAP-PHP.B3
FEATURES             Location/Qualifiers
     source          1..8543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    73..91
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    110..128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    147..165
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    184..202
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    221..239
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    258..276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    295..313
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3030..3041
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     rep_origin      complement(3597..4185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3677..3696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4356..5147,5148..5216))
                     /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     4979..4998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5217..5321)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5451..5473
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5465..5482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5466..5482
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5492..5511
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5492..5510
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8100..8210
                     /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(8249..8267)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8250..8267)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8287..8304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8293..8517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ggctcgaatc aagctatcaa gtgccacctg acgtctccct atcagtgata gagaagtcga
       61 cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct ccctatcagt
      121 gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta cgtctagaac
      181 gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag tgatagagaa
      241 ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta
      301 tcagtgatag agaaggtacc ccctatataa gcagagagat ctgttcaaat ttgaactgac
      361 taagcggctc ccgccagatt ttggcaagat tactaagcag gaagtcaagg acttttttgc
      421 ttgggcaaag gtcaatcagg tgccggtgac tcacgagttt aaagttccca gggaattggc
      481 gggaactaaa ggggcggaga aatctctaaa acgcccactg ggtgacgtca ccaatactag
      541 ctataaaagt ctggagaagc gggccaggct ctcatttgtt cccgagacgc ctcgcagttc
      601 agacgtgact gttgatcccg ctcctctgcg accgctagct tcgatcaact acgcagacag
      661 gtaccaaaac aagtgttctc gtcacgtggg cattaatctg attctgtttc cctgcagaca
      721 atgcgagaga atgaatcaga actcaaatat ctgcttcact cacggacaga aagactgttt
      781 agagtgcttt cccgtgtcag aatctcaacc cgtttctgtc gtcaaaaagg cgtatcagaa
      841 actgtgctac attcatcata tcatgggaaa ggtgccagac gcttgcactg cctgcgatct
      901 ggtcaatgtg gatttggatg actgcatctt tgaacaataa atgacttaag ccaggtatgg
      961 ctgccgatgg ttatcttcca gattggctcg aggacaacct tagtgaagga attcgcgagt
     1021 ggtgggcttt gaaacctgga gcccctcaac ccaaggcaaa tcaacaacat caagacaacg
     1081 ctagaggtct tgtgcttccg ggttacaaat accttggacc cggcaacgga ctcgacaagg
     1141 gggagccggt caacgcagca gacgcggcgg ccctcgagca cgacaaagcc tacgaccagc
     1201 agctcaaggc cggagacaac ccgtacctca agtacaacca cgccgacgcc gagttccagg
     1261 agcggctcaa agaagatacg tcttttgggg gcaacctcgg gcgagcagtc ttccaggcca
     1321 aaaagaggct tcttgaacct cttggtctgg ttgaggaagc ggctaagacg gctcctggaa
     1381 agaagaggcc tgtagagcag tctcctcagg aaccggactc ctccgcgggt attggcaaat
     1441 cgggtgcaca gcccgctaaa aagagactca atttcggtca gactggcgac acagagtcag
     1501 tcccagaccc tcaaccaatc ggagaacctc ccgcagcccc ctcaggtgtg ggatctctta
     1561 caatggcttc aggtggtggc gcaccagtgg cagacaataa cgaaggtgcc gatggagtgg
     1621 gtagttcctc gggaaattgg cattgcgatt cccaatggct gggggacaga gtcatcacca
     1681 ccagcacccg aacctgggcc ctgcccacct acaacaatca cctctacaag caaatctcca
     1741 acagcacatc tggaggatct tcaaatgaca acgcctactt cggctacagc accccctggg
     1801 ggtattttga cttcaacaga ttccactgcc acttctcacc acgtgactgg cagcgactca
     1861 tcaacaacaa ctggggattc cggcctaagc gactcaactt caagctcttt aacattcagg
     1921 tcaaagaggt tacggacaac aatggagtca agaccatcgc caataacctt accagcacgg
     1981 tccaggtctt cacggactca gactatcagc tcccgtacgt gctcgggtcg gctcacgagg
     2041 gctgcctccc gccgttccca gcggacgttt tcatgattcc tcagtacggg tatctgacgc
     2101 ttaatgatgg aagccaggcc gtgggtcgtt cgtcctttta ctgcctggaa tatttcccgt
     2161 cgcaaatgct aagaacgggt aacaacttcc agttcagcta cgagtttgag aacgtacctt
     2221 tccatagcag ctacgctcac agccaaagcc tggaccgact aatgaatcca ctcatcgacc
     2281 aatacttgta ctatctctct agaactatta acggttctgg acagaatcaa caaacgctaa
     2341 aattcagtgt ggccggaccc agcaacatgg ctgtccaggg aagaaactac atacctggac
     2401 ccagctaccg acaacaacgt gtctcaacca ctgtgactca aaacaacaac agcgaatttg
     2461 cttggcctgg agcttcttct tgggctctca atggacgtaa tagcttgatg aatcctggac
     2521 ctgctatggc ctctcacaaa gaaggagagg accgtttctt tcctttgtct ggatctttaa
     2581 tttttggcaa acaaggtact ggcagagaca acgtggatgc ggacaaagtc atgataacca
     2641 acgaagaaga aattaaaact actaacccgg tagcaacgga gtcctatgga caagtggcca
     2701 caaaccacca gagtgcccaa tttacgttga cgacgcctaa ggcacaggcg cagaccggtt
     2761 gggttcaaaa ccaaggaata cttccgggta tggtttggca ggacagagat gtgtacctgc
     2821 aaggacccat ttgggccaaa attcctcaca cggacggcaa ctttcaccct tctccgctga
     2881 tgggagggtt tggaatgaag cacccgcctc ctcagatcct catcaaaaac acacctgtac
     2941 ctgcggatcc tccaacggcc ttcaacaagg acaagctgaa ctctttcatc acccagtatt
     3001 ctactggtca agtcagcgtg gagatcgagt gggagctgca gaaggaaaac agcaagcgct
     3061 ggaacccgga gatccagtac acttccaact attacaagtc taataatgtt gaatttgctg
     3121 ttaatactga aggtgtatat agtgaacccc gccccattgg caccagatac ctgactcgta
     3181 atctgtaagt cgacttgctt gttaatcaat aaaccgttta attcgtttca gttgaacttt
     3241 ggtctctgcg aagggcaatt cgtttaaacc tgcaggacta gaggtcctgt attagaggtc
     3301 acgtgagtgt tttgcgacat tttgcgacac catgtggtca cgctgggtat ttaagcccga
     3361 gtgagcacgc agggtctcca ttttgaagcg ggaggtttga acgcgcagcc gccaagccga
     3421 attctgcaga tatcacatgt cctaggaact atcgatccat cacactggcg gccgctcgac
     3481 tagagcggcc gccaccgcgg tggagctcca gcttttgcgg accgaatcgg aaagaacatg
     3541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4021 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4261 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4321 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4381 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4441 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4501 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4561 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4621 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4681 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4741 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4801 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4861 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4921 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4981 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5041 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5101 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5161 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5221 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5281 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5341 ccacctgacg tcccaatgat acgcgtcggt gcgggcctct tcgctattac gccagctggc
     5401 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     5461 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     5521 tacatcgacg gtatcggggg agctcgcagg gtctccattt tgaagcggga ggtttgaacg
     5581 cgcagccgcc atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga
     5641 gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt
     5701 gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga
     5761 gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggctct
     5821 tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac
     5881 caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat
     5941 tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac
     6001 cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt
     6061 gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag
     6121 cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc
     6181 gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag
     6241 atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac
     6301 ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc
     6361 caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac
     6421 taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg
     6481 gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct
     6541 gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac
     6601 taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt
     6661 aaactggacc aatgagaact ttcccttcaa cgactgtgtg gacaagatgg tgatctggtg
     6721 ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag
     6781 caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat
     6841 cgtcacctcc aacaccaata tgtgcgccgt gattgacggg aactcaacga ccttcgaaca
     6901 ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga
     6961 ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt
     7021 ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc
     7081 cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac
     7141 gtcagacgcg gaagcttcga tcaactacgc ggacaggtac caaaacaaat gttctcgtca
     7201 cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagactga atcagaattc
     7261 aaatatctgc ttcactcacg gtgtcaaaga ctgtttagag tgctttcccg tgtcagaatc
     7321 tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcacatcat
     7381 gggaaaggtg ccagacgctt gcactgcttg cgacctggtc aatgtggact tggatgactg
     7441 tgtttctgaa caataaatga cttaaaccag gtatgagtcg gctggataaa tctaaagtca
     7501 taaacggcgc tctggaatta ctcaatgaag tcggtatcga aggcctgacg acaaggaaac
     7561 tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg
     7621 ccctgctcga tgccctggcc atcgagatgc tggacaggca tcatacccac ttctgccccc
     7681 tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc
     7741 tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt
     7801 acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg
     7861 cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg
     7921 agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc
     7981 tgagacaagc aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc
     8041 tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg
     8101 acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc
     8161 ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaaa
     8221 tgcatgaatt cgatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     8281 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     8341 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     8401 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     8461 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     8521 ctgaggcgga aagaaccagc tgg
//
