LOCUS       sequence-329421-        7401 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 7401)
  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 7401)
  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..7401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   75..215
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   86..215
                     /label=AAV2 ITR
     protein_bind    join(372..390,391..413,414..432,433..455,456..474,475..497,
                     498..516,517..539,540..558,559..581,582..600,601..623,
                     624..642)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        675..713
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    complement(848..881)
                     /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(892..925)
                     /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(2525..2590)
                     /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(2600..3304)
                     /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(2875..2894)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3144..3162
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3739..3748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3789..3822
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3833..3866
                     /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    3890..4478
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3943..3963)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4361..4372)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4513..4530)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4519..4726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4749..4878
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4940..5528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5020..5039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5699..6490,6491..6559))
                     /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     6322..6341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6560..6664)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6732..6750
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6788..6810)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6910..6929
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6946..7401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7083..7104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7295..7314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
       61 ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
      121 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      181 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgaa acaattctgc
      241 aggaattcgg ggccgcggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag
      301 cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct tatataggtc
      361 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
      421 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
      481 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
      541 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
      601 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagctc
      661 ggtacccggg tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggccccgaat tcgagctcgg tacccgggga tcctctagtc
      841 gacgtcgata acttcgtata gcatacatta tacgaagtta tccgaatcgc aataacttcg
      901 tataggatac tttatacgaa gttatttggc gcgtcgaggc aagctttcat tagagccgtg
      961 tctcgccccc agatttatga ctttcccaac tgctgatgat tttaccgctc tggggtgtga
     1021 cagacacttc tctaccggtc cccctcagat tgtgctgagt tggctcggag cggttcactc
     1081 gtcgccagca ggtcatcagg aaaatgatca gcatgagagc agtcagtgct ccagcggaga
     1141 gcagcacgta cttgccccaa ttaggcagtc cgagatcgac gccactgatc cgttcgtgca
     1201 cgtcgggcag gtggacctcc acaaaatctt cggcctcgtc gccgtttttg aacactgtgc
     1261 ttgggtctgc gagaggatgc atcaggggaa tgacgcttga cacgagcagt tccatgtgct
     1321 gctggagcag actagactgc atctcgggga tcagcacgtt accatctggc cccagaatga
     1381 taccattaaa gaagaccccg ttcacatgtg ggtgacatcg acccccgact cgcaggcatc
     1441 ccttactagg aatgatttca ttccaggttc tcacagattt ataatgggcg tctgcctcca
     1501 tcagtgtctt gttaaagatg gtgtatgctt ttccgaagcc agggacgagc ttcctcaggt
     1561 ggctcagcct cctgaagctc acgctcttgg tagtcataat tgactccaga gcatcgagac
     1621 attcctccct tttcttcacg agttcctcga ccaccagatg ttcgatctca tcgcttcgga
     1681 agtcgtgcag attcacgagc tgcccagggg ggcaccactt agtctcgtta gatgtctgca
     1741 tagcgaccca ggttccgtcc atcagccgga gtcccagcac gccacagagt ttcagcttgc
     1801 aggcaccttt gaggctctta tacagccctc tttcatccac aaatccacaa gtctcgctgc
     1861 cttttgaggc cctcttgccc ctgctgttgg tgaagatgtc gcaggacatc cccagccttg
     1921 gattctcagg catccagatt gtataatcat ggttggtgga acagtaagtg gagctgacgg
     1981 ccactccgga gcagtttccg ccgggaaaca ctggactgtg gagagatcgg tcgtaggggt
     2041 ccaggtcggc gacagatggg gaaatgatga ccaggctctc ttttgtggtc ttcacagtgc
     2101 gcagccaatg atagtctggg taaggattgt gcaggctttc ctcatagcgg gggtcgccag
     2161 ccatcttcca gttgtaagct gcccgacagg cgtctggtgt tggccgaaag tgttttctct
     2221 tgaaagttgt ggtgacatat cccacaaaat ttgtgtaggt ttctgcctcg gtgaccacac
     2281 cagtgcatgt gaacccgttc atcttaatag cagagatata gcccactttc agctccatgt
     2341 aggagaatcc gctcaggttg gtacagcctt cgtcctcgac cacgagattg ttagggcatg
     2401 acagatggtg aatatcgata ggactccagg ggcccagctt gtcggggata gtgtagatag
     2461 ggaatttacc aaagcagagt gggaagacga gcagagggac gaagaggaga gcctgtggga
     2521 ccatgggccc tgggttggac tccacgtctc ccgccaactt gagaaggtca aaattcaaag
     2581 tctgtttcac tccggatccc ttgtacagct cgtccatgcc gccggtggag tggcggccct
     2641 cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccaact
     2701 tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg taggtggtct
     2761 tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg atctcgccct
     2821 tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc atggtcttct
     2881 tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga
     2941 actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg tcctcgaagt
     3001 tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt
     3061 cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac aggatgtccc
     3121 aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg gtgccctcgt
     3181 aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca
     3241 tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc tcgcccttgc
     3301 tcaccccacc ccccttggat gcgctttcaa gactgaagat gtcagacctg ctttttgctt
     3361 tggacttccc ccatgcagcg ataccaccca cagctaccag gcagcacagg agcactgcag
     3421 tgatcagcat ccacatgcgg ccgtgattcc tggctggcag agcacgtcca ggagcagggg
     3481 cagtgggagc ctctgtgccg ttgtccgtgg gcaccgcggg gatcgcgctg gtcccgcagc
     3541 cccactcgtc ccgcccgtcg tcgcagtcgg ggtgtccgtc gcacagccag tcctgcgggt
     3601 aacactcccc gtgggcaccg ggcggctccg agcagcggaa ctgaccgggg gggcaacggg
     3661 acaaagaacc gttaccggac ccgttaccgg tcacgttacc gggcagcagc agcagcagca
     3721 gcgcgggcag cagccgcgcc atggtggcga attgctagct agttggcgcg ccaacctggc
     3781 cggccttgat aacttcgtat aatgtatgct atacgaagtt atccgaatcg caataacttc
     3841 gtataaagta tcctatacga agttatcgaa gatcatcaag cttatcgata atcaacctct
     3901 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3961 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4021 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4081 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4141 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4201 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4261 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     4321 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4381 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4441 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     4501 gagatcgatc tgcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4561 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4621 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4681 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggacacg tgcggaccga
     4741 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4801 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag
     4861 cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4921 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4981 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5041 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5101 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5161 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5221 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5281 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5341 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5401 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5461 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5521 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5581 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5641 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5701 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5761 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5821 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5881 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5941 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6001 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6061 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6121 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6181 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6241 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6301 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6361 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6421 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6481 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6541 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6601 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6661 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6721 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     6781 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     6841 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     6901 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta
     6961 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga
     7021 ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg
     7081 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     7141 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg
     7201 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     7261 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     7321 ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     7381 atgcgccgct acagggcgcg t
//
