LOCUS Exported 7073 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of hM4Di-mCherry in a cre-dependent fashion. ACCESSION . VERSION . KEYWORDS pAAV hSyn FLEx-loxp hM4Di-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7073) AUTHORS Beier KT, Kim CK, Hoerbelt P, Hung LW, Heifets BD, DeLoach KE, Mosca TJ, Neuner S, Deisseroth K, Luo L, Malenka RC TITLE Rabies screen reveals GPe control of cocaine-triggered plasticity. JOURNAL Nature. 2017 Sep 21;549(7672):345-350. doi: 10.1038/nature23888. Epub 2017 Sep 13. PUBMED 28902833 REFERENCE 2 (bases 1 to 7073) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7073 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 1..10 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 1459..1468 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1465..2175 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1610..1628) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1878..1897 /label=mCherry-F /note="mCherry, forward primer" protein_bind 2182..2215 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 2278..2311 /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)." promoter complement(2387..2834) /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" repeat_region 2879..3008 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" rep_origin complement(3070..3658) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3150..3169) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3829..4689) /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 4452..4471 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4690..4794) /gene="bla" /label=AmpR promoter primer_bind 4862..4880 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4918..4940) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5040..5059 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(5076..5531) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5213..5234) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5425..5444 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 5606..5746 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5617..5746 /label=AAV2 ITR polyA_signal 5786..6262 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind 6044..6063 /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" misc_feature 6294..6882 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 6400..6411 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 6809..6829 /label=WPRE-R /note="WPRE, reverse primer" protein_bind complement(6930..6963) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(7034..7067) /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)." ORIGIN 1 gccgccatgg ccaacttcac acctgtcaat ggcagctcgg gcaatcagtc cgtgcgcctg 61 gtcacgtcat catcccacaa tcgctatgag acggtggaaa tggtcttcat tgccacagtg 121 acaggctccc tgagcctggt gactgtcgtg ggcaacatcc tggtgatgct gtccatcaag 181 gtcaacaggc agctgcagac agtcaacaac tacttcctct tcagcctggc gtgtgctgat 241 ctcatcatag gcgccttctc catgaacctc tacaccgtgt acatcatcaa gggctactgg 301 cccctgggcg ccgtggtctg cgacctgtgg ctggccctgg actgcgtggt gagcaacgcc 361 tccgtcatga accttctcat catcagcttt gaccgctact tctgcgtcac caagcctctc 421 acctaccctg cccggcgcac caccaagatg gcaggcctca tgattgctgc tgcctgggta 481 ctgtccttcg tgctctgggc gcctgccatc ttgttctggc agtttgtggt gggtaagcgg 541 acggtgcccg acaaccagtg cttcatccag ttcctgtcca acccagcagt gacctttggc 601 acagccattg ctggcttcta cctgcctgtg gtcatcatga cggtgctgta catccacatc 661 tccctggcca gtcgcagccg agtccacaag caccggcccg agggcccgaa ggagaagaaa 721 gccaagacgc tggccttcct caagagccca ctaatgaagc agagcgtcaa gaagcccccg 781 cccggggagg ccgcccggga ggagctgcgc aatggcaagc tggaggaggc ccccccgcca 841 gcgctgccac cgccaccgcg ccccgtggct gataaggaca cttccaatga gtccagctca 901 ggcagtgcca cccagaacac caaggaacgc ccagccacag agctgtccac cacagaggcc 961 accacgcccg ccatgcccgc ccctcccctg cagccgcggg ccctcaaccc agcctccaga 1021 tggtccaaga tccagattgt gacgaagcag acaggcaatg agtgtgtgac agccattgag 1081 attgtgcctg ccacgccggc tggcatgcgc cctgcggcca acgtggcccg caagttcgcc 1141 agcatcgctc gcaaccaggt gcgcaagaag cggcagatgg cggcccggga gcgcaaagtg 1201 acacgaacga tctttgccat tctgctggcc ttcatcctca cctggacgcc ctacaacgtc 1261 atggtcctgg tgaacacctt ctgccagagc tgcatccctg acacggtgtg gtccattggc 1321 tactggctct gctacgtcaa cagcaccatc aaccctgcct gctatgctct gtgcaacgcc 1381 acctttaaaa agaccttccg gcacctgctg ctgtgccagt atcggaacat cggcactgcc 1441 aggcgggatc caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc 1501 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc 1561 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag 1621 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac 1681 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc 1741 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg 1801 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc 1861 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc 1921 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg 1981 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg 2041 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac 2101 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac 2161 gagctgtaca agtaagtcga cataacttcg tatagcatac attatacgaa gttatgtgtc 2221 gatggtgatg cttggcaatt cgggtcgatg gtgaagcatt ggtcttccta ttttgatata 2281 acttcgtata ggatacttta tacgaagtta tcccggccgg gcccgcggta tcaaatcaat 2341 tctttgccaa aatgatgaga cagcacagga tccggtacct tctcgactgc gctctcaggc 2401 acgacacgac tcctccgctg cccaccgcag actgaggcag cgctgagtcg ccggcgccgc 2461 agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc ggtccggctg 2521 ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag gggagtttgc gggggaccgg 2581 cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg gcgcgcgccg ccaggcgggg 2641 gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca cgcgcctcgc cgcatcctgt 2701 ttcccctccc cctctctgat aggggatgcg caatttgggg aatgggggtt gggtgcttgt 2761 ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc ccgcctcatc ctggtcctaa 2821 aacccacttg cactcatacg cagggccctc tgcagtctag acacacgcgt gcggccgcag 2881 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc 2941 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg 3001 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 3061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 3121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 3181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 3241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 3301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 3361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 3421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 3481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 3541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 3601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 3661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 3721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 3781 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 3841 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 3901 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 3961 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 4021 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 4081 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 4141 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 4201 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 4261 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 4321 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 4381 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 4441 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 4501 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 4561 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 4621 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 4681 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 4741 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 4801 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 4861 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 4921 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 4981 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 5041 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt 5101 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg 5161 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg 5221 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 5281 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg 5341 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 5401 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct 5461 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct 5521 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta 5581 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag 5641 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag 5701 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc 5761 cgcacgtggt tacctacaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta 5821 atcccagcaa tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca 5881 gcctggccaa tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg 5941 tcatgcatgc ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc 6001 aggaggcgga gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat 6061 agaccccgca ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc 6121 tccaccccat aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta 6181 ttaggacaag gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg 6241 aggggtcaca gggatgccac ccgtagatct ctcgagcagc gctcggtatc gatgcgggga 6301 ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg 6361 cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta 6421 gcagaaggac gtcccgcgca gaatccaggt ggcaacatag gcgagcagcc aaggaaagga 6481 cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca 6541 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa 6601 agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc 6661 aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga 6721 cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg 6781 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat 6841 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa gcttgatatc 6901 gaattcatcg atcggccgca tataagctta taacttcgta taatgtatgc tatacgaagt 6961 tatgggtcga tggtgagatc tggactagag ggtcgatggt gatgcttgga tcaaaatagg 7021 aagaatccca atgataactt cgtataaagt atcctatacg aagttatagg cgc //