LOCUS       sequence-246392-        6185 bp    DNA     circular SYN 14-APR-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 6185)
  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 6185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..583
                     /label=chicken beta-actin promoter
     primer_bind     913..935
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     997..1016
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1065..1098)
                     /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(1149..1182)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1206..1316)
                     /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    1969..2002
                     /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    2053..2086
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2103..2691
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2156..2176)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2574..2585)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2723..3199
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2922..2941)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3239..3379
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3239..3368
                     /label=AAV2 ITR
     rep_origin      3454..3909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3541..3560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3751..3772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3926..3945)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4045..4067
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4105..4123)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4191..4295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4296..4364,4365..5156)
                     /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(4514..4533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5327..5915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5816..5835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5977..6106
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      421 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
      481 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      541 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      601 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      661 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      721 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      781 ggagcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
      841 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
      901 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
      961 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1021 ctcatcattt tggcaaagaa ttgcggccca acggtacctc cggaataact tcgtatagga
     1081 tactttatac gaagttatgc agaatggtag ctggattgta gctgctatta gcaatatgaa
     1141 acctcttaat aacttcgtat agcatacatt atacgaagtt atggcgcgcc aagcttttac
     1201 ccggggagca tgtcaaggtc aaaatcgtca agagcgtcag caggcagcat atcaaggtca
     1261 aagtcgtcaa gggcatcggc tgggagcatg tctaagtcaa aatcgtcaag ggcgtcggcc
     1321 ggcccgccgc tttcgcactt tagctgtttc tccaggccac atatgattag ttccaggccg
     1381 aaaaggaagg caggttcggc tccctgccgg tcgaacagct caattgcttg tctcagaagt
     1441 gggggcatag aatcggtggt aggtgtctct ctttcctctt ttgctacttg atgctcctgt
     1501 tcctccaata cgcagcccag tgtaaagtgg cccacggcgg acagagcgta cagtgcgttc
     1561 tccagggaga agccttgctg acacaggaac gcgagctgat tttccagggt ttcgtactgt
     1621 ttctctgttg ggcgggtgcc gagatgcact ttagccccgt cgcgatgtga gaggagagca
     1681 cagcggaatg acttggcgtt gttccgcaga aagtcttgcc atgactcgcc ttccaggggg
     1741 cagaagtggg tatgatgcct gtccagcatc tcgatggcca gggcatcgag cagggcccgc
     1801 ttgttcttca cgtgccagta cagggtaggc tgctcaactc ccagcttttg agcgagtttc
     1861 cttgtcgtca ggccttcgat accgacttca ttgagtaatt ccagagcgcc gtttatgact
     1921 ttagatttat ccagccgact catggtggcg gatatcggat ccgctagcat aacttcgtat
     1981 aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt attctttaga
     2041 atggtgcaaa gaataacttc gtataatgta tgctatacga agttatgaat tcgcttatcg
     2101 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2161 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2221 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2281 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2341 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2401 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2461 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2521 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2581 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2641 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     2701 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     2761 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2821 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     2881 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     2941 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     3001 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     3061 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     3121 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     3181 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     3241 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3301 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3361 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3421 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3481 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3541 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3601 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3661 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3721 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3781 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3841 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3901 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3961 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4021 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4081 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4141 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4201 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4261 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4321 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4381 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4441 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4501 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4561 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4621 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4681 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4741 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4801 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4861 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4921 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4981 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5041 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5101 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5161 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5221 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5281 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5341 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5401 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5461 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5521 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5581 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5641 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5701 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5761 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5821 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5881 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5941 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     6001 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     6061 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     6121 aacactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6181 ttccg
//
