LOCUS       sequence-250685-        8469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-standard AAV2 rep-AAV-PHP.V2 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 8469)
  AUTHORS   Ravindra Kumar S, Miles TF, Chen X, Brown D, Dobreva T, Huang Q, 
            Ding X, Luo Y, Einarsson PH, Greenbaum A, Jang MJ, Deverman BE, 
            Gradinaru V
  TITLE     Multiplexed Cre-dependent selection yields systemic AAVs for 
            targeting distinct brain cell types.
  JOURNAL   Nat Methods. 2020 May;17(5):541-550. doi: 10.1038/s41592-020-0799-7.
            Epub 2020 Apr 20.
  PUBMED    32313222
REFERENCE   2  (bases 1 to 8469)
  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/s41592-020-0799-7"; journalName: "Nat Methods"; date: 
            "2020-05"; volume: "17"; issue: "5"; pages: "541-550"
COMMENT     Sequence Label: pUCmini-iCAP-PHP.V2
FEATURES             Location/Qualifiers
     source          1..8469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..121
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     113..130
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     114..130
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     140..159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        140..158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2748..2858
                     /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(2897..2915)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2898..2915)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2935..2952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2941..3165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3227..3245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3264..3282
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3301..3319
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3338..3356
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3375..3393
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3412..3430
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             6147..6158
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     rep_origin      complement(6714..7302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6794..6813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7473..8264,8265..8333))
                     /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     8096..8115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8334..8438)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgatac gcgtcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
       61 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      121 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta catcgacggt
      181 atcgggggag ctcgcagggt ctccattttg aagcgggagg tttgaacgcg cagccgccat
      241 gccggggttt tacgagattg tgattaaggt ccccagcgac cttgacgagc atctgcccgg
      301 catttctgac agctttgtga actgggtggc cgagaaggaa tgggagttgc cgccagattc
      361 tgacatggat ctgaatctga ttgagcaggc acccctgacc gtggccgaga agctgcagcg
      421 cgactttctg acggaatggc gccgtgtgag taaggccccg gaggctcttt tctttgtgca
      481 atttgagaag ggagagagct acttccacat gcacgtgctc gtggaaacca ccggggtgaa
      541 atccatggtt ttgggacgtt tcctgagtca gattcgcgaa aaactgattc agagaattta
      601 ccgcgggatc gagccgactt tgccaaactg gttcgcggtc acaaagacca gaaatggcgc
      661 cggaggcggg aacaaggtgg tggatgagtg ctacatcccc aattacttgc tccccaaaac
      721 ccagcctgag ctccagtggg cgtggactaa tatggaacag tatttaagcg cctgtttgaa
      781 tctcacggag cgtaaacggt tggtggcgca gcatctgacg cacgtgtcgc agacgcagga
      841 gcagaacaaa gagaatcaga atcccaattc tgatgcgccg gtgatcagat caaaaacttc
      901 agccaggtac atggagctgg tcgggtggct cgtggacaag gggattacct cggagaagca
      961 gtggatccag gaggaccagg cctcatacat ctccttcaat gcggcctcca actcgcggtc
     1021 ccaaatcaag gctgccttgg acaatgcggg aaagattatg agcctgacta aaaccgcccc
     1081 cgactacctg gtgggccagc agcccgtgga ggacatttcc agcaatcgga tttataaaat
     1141 tttggaacta aacgggtacg atccccaata tgcggcttcc gtctttctgg gatgggccac
     1201 gaaaaagttc ggcaagagga acaccatctg gctgtttggg cctgcaacta ccgggaagac
     1261 caacatcgcg gaggccatag cccacactgt gcccttctac gggtgcgtaa actggaccaa
     1321 tgagaacttt cccttcaacg actgtgtgga caagatggtg atctggtggg aggaggggaa
     1381 gatgaccgcc aaggtcgtgg agtcggccaa agccattctc ggaggaagca aggtgcgcgt
     1441 ggaccagaaa tgcaagtcct cggcccagat agacccgact cccgtgatcg tcacctccaa
     1501 caccaatatg tgcgccgtga ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt
     1561 gcaagaccgg atgttcaaat ttgaactcac ccgccgtctg gatcatgact ttgggaaggt
     1621 caccaagcag gaagtcaaag actttttccg gtgggcaaag gatcacgtgg ttgaggtgga
     1681 gcatgaattc tacgtcaaaa agggtggagc caagaaaaga cccgccccca gtgacgcaga
     1741 tataagtgag cccaaacggg tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga
     1801 agcttcgatc aactacgcgg acaggtacca aaacaaatgt tctcgtcacg tgggcatgaa
     1861 tctgatgctg tttccctgca gacaatgcga gagactgaat cagaattcaa atatctgctt
     1921 cactcacggt gtcaaagact gtttagagtg ctttcccgtg tcagaatctc aacccgtttc
     1981 tgtcgtcaaa aaggcgtatc agaaactgtg ctacattcat cacatcatgg gaaaggtgcc
     2041 agacgcttgc actgcttgcg acctggtcaa tgtggacttg gatgactgtg tttctgaaca
     2101 ataaatgact taaaccaggt atgagtcggc tggataaatc taaagtcata aacggcgctc
     2161 tggaattact caatgaagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
     2221 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
     2281 ccctggccat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt
     2341 catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc
     2401 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
     2461 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
     2521 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
     2581 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa
     2641 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
     2701 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg
     2761 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     2821 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaatg catgaattcg
     2881 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     2941 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3001 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3061 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3121 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3181 gaaccagctg gggctcgaat caagctatca agtgccacct gacgtctccc tatcagtgat
     3241 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc
     3301 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
     3361 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca
     3421 gtgatagaga aggtaccccc tatataagca gagagatctg ttcaaatttg aactgactaa
     3481 gcggctcccg ccagattttg gcaagattac taagcaggaa gtcaaggact tttttgcttg
     3541 ggcaaaggtc aatcaggtgc cggtgactca cgagtttaaa gttcccaggg aattggcggg
     3601 aactaaaggg gcggagaaat ctctaaaacg cccactgggt gacgtcacca atactagcta
     3661 taaaagtctg gagaagcggg ccaggctctc atttgttccc gagacgcctc gcagttcaga
     3721 cgtgactgtt gatcccgctc ctctgcgacc gctagcttcg atcaactacg cagacaggta
     3781 ccaaaacaag tgttctcgtc acgtgggcat taatctgatt ctgtttccct gcagacaatg
     3841 cgagagaatg aatcagaact caaatatctg cttcactcac ggacagaaag actgtttaga
     3901 gtgctttccc gtgtcagaat ctcaacccgt ttctgtcgtc aaaaaggcgt atcagaaact
     3961 gtgctacatt catcatatca tgggaaaggt gccagacgct tgcactgcct gcgatctggt
     4021 caatgtggat ttggatgact gcatctttga acaataaatg acttaagcca ggtatggctg
     4081 ccgatggtta tcttccagat tggctcgagg acaaccttag tgaaggaatt cgcgagtggt
     4141 gggctttgaa acctggagcc cctcaaccca aggcaaatca acaacatcaa gacaacgcta
     4201 gaggtcttgt gcttccgggt tacaaatacc ttggacccgg caacggactc gacaaggggg
     4261 agccggtcaa cgcagcagac gcggcggccc tcgagcacga caaagcctac gaccagcagc
     4321 tcaaggccgg agacaacccg tacctcaagt acaaccacgc cgacgccgag ttccaggagc
     4381 ggctcaaaga agatacgtct tttgggggca acctcgggcg agcagtcttc caggccaaaa
     4441 agaggcttct tgaacctctt ggtctggttg aggaagcggc taagacggct cctggaaaga
     4501 agaggcctgt agagcagtct cctcaggaac cggactcctc cgcgggtatt ggcaaatcgg
     4561 gtgcacagcc cgctaaaaag agactcaatt tcggtcagac tggcgacaca gagtcagtcc
     4621 cagaccctca accaatcgga gaacctcccg cagccccctc aggtgtggga tctcttacaa
     4681 tggcttcagg tggtggcgca ccagtggcag acaataacga aggtgccgat ggagtgggta
     4741 gttcctcggg aaattggcat tgcgattccc aatggctggg ggacagagtc atcaccacca
     4801 gcacccgaac ctgggccctg cccacctaca acaatcacct ctacaagcaa atctccaaca
     4861 gcacatctgg aggatcttca aatgacaacg cctacttcgg ctacagcacc ccctgggggt
     4921 attttgactt caacagattc cactgccact tctcaccacg tgactggcag cgactcatca
     4981 acaacaactg gggattccgg cctaagcgac tcaacttcaa gctctttaac attcaggtca
     5041 aagaggttac ggacaacaat ggagtcaaga ccatcgccaa taaccttacc agcacggtcc
     5101 aggtcttcac ggactcagac tatcagctcc cgtacgtgct cgggtcggct cacgagggct
     5161 gcctcccgcc gttcccagcg gacgttttca tgattcctca gtacgggtat ctgacgctta
     5221 atgatggaag ccaggccgtg ggtcgttcgt ccttttactg cctggaatat ttcccgtcgc
     5281 aaatgctaag aacgggtaac aacttccagt tcagctacga gtttgagaac gtacctttcc
     5341 atagcagcta cgctcacagc caaagcctgg accgactaat gaatccactc atcgaccaat
     5401 acttgtacta tctctctaga actattaacg gttctggaca gaatcaacaa acgctaaaat
     5461 tcagtgtggc cggacccagc aacatggctg tccagggaag aaactacata cctggaccca
     5521 gctaccgaca acaacgtgtc tcaaccactg tgactcaaaa caacaacagc gaatttgctt
     5581 ggcctggagc ttcttcttgg gctctcaatg gacgtaatag cttgatgaat cctggacctg
     5641 ctatggcctc tcacaaagaa ggagaggacc gtttctttcc tttgtctgga tctttaattt
     5701 ttggcaaaca aggtactggc agagacaacg tggatgcgga caaagtcatg ataaccaacg
     5761 aagaagaaat taaaactact aacccggtag caacggagtc ctatggacaa gtggccacaa
     5821 accaccagag tgcccaaacc accctcaaac ccttcctcgc acaggcgcag accggttggg
     5881 ttcaaaacca aggaatactt ccgggtatgg tttggcagga cagagatgtg tacctgcaag
     5941 gacccatttg ggccaaaatt cctcacacgg acggcaactt tcacccttct ccgctgatgg
     6001 gagggtttgg aatgaagcac ccgcctcctc agatcctcat caaaaacaca cctgtacctg
     6061 cggatcctcc aacggccttc aacaaggaca agctgaactc tttcatcacc cagtattcta
     6121 ctggtcaagt cagcgtggag atcgagtggg agctgcagaa ggaaaacagc aagcgctgga
     6181 acccggagat ccagtacact tccaactatt acaagtctaa taatgttgaa tttgctgtta
     6241 atactgaagg tgtatatagt gaaccccgcc ccattggcac cagatacctg actcgtaatc
     6301 tgtaagtcga cttgcttgtt aatcaataaa ccgtttaatt cgtttcagtt gaactttggt
     6361 ctctgcgaag ggcaattcgt ttaaacctgc aggactagag gtcctgtatt agaggtcacg
     6421 tgagtgtttt gcgacatttt gcgacaccat gtggtcacgc tgggtattta agcccgagtg
     6481 agcacgcagg gtctccattt tgaagcggga ggtttgaacg cgcagccgcc aagccgaatt
     6541 ctgcagatat cacatgtcct aggaactatc gatccatcac actggcggcc gctcgactag
     6601 agcggccgcc accgcggtgg agctccagct tttgcggacc gaatcggaaa gaacatgtga
     6661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8461 cctgacgtc
//
