LOCUS       sequence-130617-        7238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependant TVA receptor-mCherry fusion with a mutation (66T) in 
            TVA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7238)
  AUTHORS   Miyamichi K, Shlomai-Fuchs Y, Shu M, Weissbourd BC, Luo L, Mizrahi A
  TITLE     Dissecting local circuits: parvalbumin interneurons underlie broad 
            feedback control of olfactory bulb output.
  JOURNAL   Neuron. 2013 Dec 4;80(5):1232-45. doi: 10.1016/j.neuron.2013.08.027.
            Epub 2013 Nov 14.
  PUBMED    24239125
REFERENCE   2  (bases 1 to 7238)
  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: 
            "10.1016/j.neuron.2013.08.027"; journalName: "Neuron"; date: 
            "2013-12-4- 4"; volume: "80"; issue: "5"; pages: "1232-45"
COMMENT     Sequence Label: CAG-Flex-TC66T
FEATURES             Location/Qualifiers
     source          1..7238
                     /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        231..534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        536..813
                     /label=chicken beta-actin promoter
     intron          815..1832
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1756..1778
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1840..1859
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1912..1945
                     /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    2008..2041
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2048..2755)
                     /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(2326..2345)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2595..2613
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3190..3199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3206..3239)
                     /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(3310..3343)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3367..3955
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3420..3440)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3838..3849)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3979..4455
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4178..4197)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4501..4641
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4501..4630
                     /label=AAV2 ITR
     rep_origin      4716..5171
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4803..4822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5013..5034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5188..5207)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5307..5329
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5367..5385)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5453..5557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5558..5626,5627..6418)
                     /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(5776..5795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6589..7177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7078..7097
                     /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 cgcgtaattc ccggggaatc tagttattaa
      181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
      541 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
      601 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
      661 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      721 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      781 cggccctata aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc
      841 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      901 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      961 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     1021 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     1081 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     1141 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     1201 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     1261 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     1321 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     1381 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     1441 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     1501 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     1561 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     1621 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     1681 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     1741 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     1801 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     1861 tgtctcatca ttttggcaaa gaattgattt gataccgcgg gcccggccgg gataacttcg
     1921 tataaagtat cctatacgaa gttatatcaa aataggaaga ccaatgcttc accatcgacc
     1981 cgaattgcca agcatcacca tcgacacata acttcgtata atgtatgcta tacgaagtta
     2041 tgtcgactta cttgtacagc tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt
     2101 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga
     2161 cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcag
     2221 cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc
     2281 cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc ttctgcatta
     2341 cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt
     2401 cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc
     2461 gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt
     2521 gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg
     2581 gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     2641 cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct
     2701 tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccccac
     2761 cccccttgga tgcgctttca agactgaaga tgtcagacct gctttttgct ttggacttcc
     2821 cccatgcagc gataccaccc acagctacca ggcagcacag gagcactgca gtgatcagca
     2881 tccacatgcg gccgtgattc ctggctggca gagcacgtcc aggagcaggg gcagtgggag
     2941 cctctgtgcc gttgtccgtg ggcaccgcgg ggatcgcgct ggtcccgcag ccccaggtgt
     3001 cccgcccgtc gtcgcagtcg gggtgtccgt cgcacagcca gtcctgcggg taacactccc
     3061 cgtgggcacc gggcggctcc gagcagcgga actgaccggg ggggcaacgg gacaaagaac
     3121 cgttaccgga cccgttaccg gtcacgttac cgggcagcag cagcagcagc agcgcgggca
     3181 gcagccgcgc catggtggcg cgcctataac ttcgtatagg atactttata cgaagttatc
     3241 attgggattc ttcctatttt gatccaagca tcaccatcga ccctctagtc cagatctcac
     3301 catcgaccca taacttcgta tagcatacat tatacgaagt tataagctta tatgcggccg
     3361 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3421 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3481 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3541 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3601 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3661 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3721 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3781 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3841 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3901 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     3961 taccgtagat cgatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     4021 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     4081 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     4141 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     4201 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     4261 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     4321 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     4381 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     4441 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgaacta gtgcggccgc
     4501 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4561 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4621 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4681 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4921 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4981 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5041 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     5101 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5161 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5221 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5281 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5341 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5401 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5521 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5581 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5641 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5701 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5761 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5821 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5881 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5941 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6001 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6061 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6121 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6181 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6241 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6301 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6361 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6421 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6481 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6541 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6721 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7201 acggttcctg gccttttgct ggccttttgc tcacatgt
//
