LOCUS Exported 7238 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-dependant TVA receptor-mCherry fusion with a mutation (66T) in TVA. ACCESSION . VERSION . KEYWORDS CAG-Flex-TC66T 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7238 /organism="synthetic DNA construct" /mol_type="other DNA" 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 complement(1912..1945) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(2008..2041) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 3206..3239 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 3310..3343 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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 5558..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 //