LOCUS       sequence-243270-        6198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome with Cre-dependent expression of tTA from the CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6198)
  AUTHORS   Rajendran PS, Challis RC, Fowlkes CC, Hanna P, Tompkins JD, Jordan 
            MC, Hiyari S, Gabris-Weber BA, Greenbaum A, Chan KY, Deverman BE, 
            Munzberg H, Ardell JL, Salama G, Gradinaru V, Shivkumar K
  TITLE     Identification of peripheral neural circuits that regulate heart 
            rate using optogenetic and viral vector strategies.
  JOURNAL   Nat Commun. 2019 Apr 26;10(1):1944. doi: 10.1038/s41467-019-09770-1.
  PUBMED    31028266
REFERENCE   2  (bases 1 to 6198)
  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 
            Commun."; date: "2019-04-26"; volume: "10(1):1944. doi"; pages: " 
            10.1038/s41467-019-09770-1"
COMMENT     Sequence Label: pAAV-CAG-DIO-tTA
FEATURES             Location/Qualifiers
     source          1..6198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..803
                     /label=chicken beta-actin promoter
     primer_bind     1133..1155
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1217..1236
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1286..1319)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1370..1403)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1427..1537)
                     /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"
     protein_bind    2190..2223
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2274..2307
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2324..2912
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2377..2397)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2795..2806)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2945..3421
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3144..3163)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3461..3601
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3461..3590
                     /label=AAV2 ITR
     rep_origin      3676..4131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3763..3782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3973..3994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4148..4167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4267..4289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4327..4345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4413..4517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4518..4586,4587..5378)
                     /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     complement(4736..4755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5549..6137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6038..6057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaacac tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      541 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      601 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg
      661 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
      721 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      781 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      841 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      901 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      961 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagcgccgg caggaaggaa
     1021 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1081 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1141 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1201 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1261 ttgcggccca acggtaccct ccggaataac ttcgtatagg atactttata cgaagttatg
     1321 cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta
     1381 tagcatacat tatacgaagt tatggcgcgc caagctttta cccggggagc atgtcaaggt
     1441 caaaatcgtc aagagcgtca gcaggcagca tatcaaggtc aaagtcgtca agggcatcgg
     1501 ctgggagcat gtctaagtca aaatcgtcaa gggcgtcggc cggcccgccg ctttcgcact
     1561 ttagctgttt ctccaggcca catatgatta gttccaggcc gaaaaggaag gcaggttcgg
     1621 ctccctgccg gtcgaacagc tcaattgctt gtctcagaag tgggggcata gaatcggtgg
     1681 taggtgtctc tctttcctct tttgctactt gatgctcctg ttcctccaat acgcagccca
     1741 gtgtaaagtg gcccacggcg gacagagcgt acagtgcgtt ctccagggag aagccttgct
     1801 gacacaggaa cgcgagctga ttttccaggg tttcgtactg tttctctgtt gggcgggtgc
     1861 cgagatgcac tttagccccg tcgcgatgtg agaggagagc acagcggaat gacttggcgt
     1921 tgttccgcag aaagtcttgc catgactcgc cttccagggg gcagaagtgg gtatgatgcc
     1981 tgtccagcat ctcgatggcc agggcatcga gcagggcccg cttgttcttc acgtgccagt
     2041 acagggtagg ctgctcaact cccagctttt gagcgagttt ccttgtcgtc aggccttcga
     2101 taccgacttc attgagtaat tccagagcgc cgtttatgac tttagattta tccagccgac
     2161 tcatggtggc ggatatcgga tccgctagca taacttcgta taaagtatcc tatacgaagt
     2221 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     2281 cgtataatgt atgctatacg aagttatgaa ttcgcttatc gataatcaac ctctggatta
     2341 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2401 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2461 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2521 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2581 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2641 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2701 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     2761 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2821 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2881 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc ctcgagagat
     2941 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     3001 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     3061 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     3121 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     3181 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     3241 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     3301 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     3361 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     3421 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3481 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3541 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3601 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3661 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3721 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3781 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3841 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3901 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3961 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4021 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4081 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4141 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4201 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4261 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4321 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4381 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgt
//
