LOCUS       sequence-323214-        7411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cre-dependent TVA-oG-mCherry under control of the TRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7411)
  AUTHORS   Sabatini PV, Wang J, Rupp AC, Affinati AH, Flak JN, Li C, Olson DP, 
            Myers MG Jr
  TITLE     tTARGIT AAVs mediate the sensitive and flexible manipulation of 
            intersectional neuronal populations in mice.
  JOURNAL   eLife. 2021 Mar 11;10. pii: 66835. doi: 10.7554/eLife.66835.
  PUBMED    33704065
REFERENCE   2  (bases 1 to 7411)
  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: "eLife."; 
            date: "2021-03-11"; volume: "10. pii: 66835. doi"; pages: " 
            10.7554/eLife.66835"
COMMENT     Sequence Label: pAAV-TRE-DIO-TVA-oG-mCherry
FEATURES             Location/Qualifiers
     source          1..7411
                     /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
     protein_bind    join(298..316,317..339,340..358,359..381,382..400,401..423,
                     424..442,443..465,466..484,485..507,508..526,527..549,
                     550..568)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        601..638
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    complement(773..806)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(817..850)
                     /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)."
     CDS             complement(2450..2515)
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             complement(2525..3229)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2800..2819)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3069..3087
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3664..3673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3714..3747
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3758..3791
                     /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)."
     misc_feature    3815..4403
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3868..3888)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4286..4297)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4438..4455)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4444..4651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4674..4814
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4674..4803
                     /label=AAV2 ITR
     rep_origin      4889..5344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4976..4995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5186..5207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5361..5380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5480..5502
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5540..5558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5626..5730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5731..5799,5800..6591)
                     /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(5949..5968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6762..7350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7251..7270
                     /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 cgcgaaacaa ttctgcagga attcggggcc
      181 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      241 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc
      301 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      361 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      421 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      481 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      541 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga
      601 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccgcggccc cgaattcgag ctcggtaccc ggggatcctc tagtcgacgt cgataacttc
      781 gtatagcata cattatacga agttatccga atcgcaataa cttcgtatag gatactttat
      841 acgaagttat ttggcgcgtc gaggcaagct ttcattagag ccgtgtctcg cccccagatt
      901 tatgactttc ccaactgctg atgattttac cgctctgggg tgtgacagac acttctctac
      961 cggtccccct cagattgtgc tgagttggct cggagcggtt cactcgtcgc cagcaggtca
     1021 tcaggaaaat gatcagcatg agagcagtca gtgctccagc ggagagcagc acgtacttgc
     1081 cccaattagg cagtccgaga tcgacgccac tgatccgttc gtgcacgtcg ggcaggtgga
     1141 cctccacaaa atcttcggcc tcgtcgccgt ttttgaacac tgtgcttggg tctgcgagag
     1201 gatgcatcag gggaatgacg cttgacacga gcagttccat gtgctgctgg agcagactag
     1261 actgcatctc ggggatcagc acgttaccat ctggccccag aatgatacca ttaaagaaga
     1321 ccccgttcac atgtgggtga catcgacccc cgactcgcag gcatccctta ctaggaatga
     1381 tttcattcca ggttctcaca gatttataat gggcgtctgc ctccatcagt gtcttgttaa
     1441 agatggtgta tgcttttccg aagccaggga cgagcttcct caggtggctc agcctcctga
     1501 agctcacgct cttggtagtc ataattgact ccagagcatc gagacattcc tcccttttct
     1561 tcacgagttc ctcgaccacc agatgttcga tctcatcgct tcggaagtcg tgcagattca
     1621 cgagctgccc aggggggcac cacttagtct cgttagatgt ctgcatagcg acccaggttc
     1681 cgtccatcag ccggagtccc agcacgccac agagtttcag cttgcaggca cctttgaggc
     1741 tcttatacag ccctctttca tccacaaatc cacaagtctc gctgcctttt gaggccctct
     1801 tgcccctgct gttggtgaag atgtcgcagg acatccccag ccttggattc tcaggcatcc
     1861 agattgtata atcatggttg gtggaacagt aagtggagct gacggccact ccggagcagt
     1921 ttccgccggg aaacactgga ctgtggagag atcggtcgta ggggtccagg tcggcgacag
     1981 atggggaaat gatgaccagg ctctcttttg tggtcttcac agtgcgcagc caatgatagt
     2041 ctgggtaagg attgtgcagg ctttcctcat agcgggggtc gccagccatc ttccagttgt
     2101 aagctgcccg acaggcgtct ggtgttggcc gaaagtgttt tctcttgaaa gttgtggtga
     2161 catatcccac aaaatttgtg taggtttctg cctcggtgac cacaccagtg catgtgaacc
     2221 cgttcatctt aatagcagag atatagccca ctttcagctc catgtaggag aatccgctca
     2281 ggttggtaca gccttcgtcc tcgaccacga gattgttagg gcatgacaga tggtgaatat
     2341 cgataggact ccaggggccc agcttgtcgg ggatagtgta gatagggaat ttaccaaagc
     2401 agagtgggaa gacgagcaga gggacgaaga ggagagcctg tgggaccatg ggccctgggt
     2461 tggactccac gtctcccgcc aacttgagaa ggtcaaaatt caaagtctgt ttcactccgg
     2521 atcccttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact
     2581 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt
     2641 aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt
     2701 agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct
     2761 cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc
     2821 cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca
     2881 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
     2941 acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca
     3001 cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg
     3061 ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     3121 cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc accttgaagc
     3181 gcatgaactc cttgatgatg gccatgttat cctcctcgcc cttgctcacc ccacccccct
     3241 tggatgcgct ttcaagactg aagatgtcag acctgctttt tgctttggac ttcccccatg
     3301 cagcgatacc acccacagct accaggcagc acaggagcac tgcagtgatc agcatccaca
     3361 tgcggccgtg attcctggct ggcagagcac gtccaggagc aggggcagtg ggagcctctg
     3421 tgccgttgtc cgtgggcacc gcggggatcg cgctggtccc gcagccccac tcgtcccgcc
     3481 cgtcgtcgca gtcggggtgt ccgtcgcaca gccagtcctg cgggtaacac tccccgtggg
     3541 caccgggcgg ctccgagcag cggaactgac cgggggggca acgggacaaa gaaccgttac
     3601 cggacccgtt accggtcacg ttaccgggca gcagcagcag cagcagcgcg ggcagcagcc
     3661 gcgccatggt ggcgaattgc tagctagttg gcgcgccaac ctggccggcc ttgataactt
     3721 cgtataatgt atgctatacg aagttatccg aatcgcaata acttcgtata aagtatccta
     3781 tacgaagtta tcgaagatca tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3841 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3901 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3961 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4021 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4081 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4141 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4201 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     4261 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4321 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4381 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat cgatctgcct
     4441 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4501 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4561 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4621 attgggaaga caatagcagg catgctgggg acacgtgcgg accgagcggc cgcaggaacc
     4681 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4741 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4801 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4861 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4921 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4981 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5041 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5101 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5161 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5221 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5281 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5341 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5401 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5461 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5521 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5581 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5701 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5761 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5821 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5881 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5941 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6001 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6061 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6121 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6181 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6241 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6301 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6361 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6421 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6481 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6541 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6601 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6901 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7381 ctggcctttt gctggccttt tgctcacatg t
//
