LOCUS Exported 6844 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Fluorescent calcium reporter and mouse vomeronasal 1 receptor 85. ACCESSION . VERSION . KEYWORDS pAAV2-GCaMP6f-P2A-Vmn1r85 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6844) AUTHORS Lee D, Kume M, Holy TE TITLE Sensory coding mechanisms revealed by optical tagging of physiologically defined neuronal types. JOURNAL Science. 2019 Dec 13;366(6471):1384-1389. doi: 10.1126/science.aax8055. PUBMED 31831669 REFERENCE 2 (bases 1 to 6844) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6844 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" enhancer 223..526 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 1056..1075 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1130..1139 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1151..1207 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 1420..1441 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1547..1568) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1808..1827) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 2399..2455 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" polyA_signal 3393..3527 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3417..3436) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3471..3490 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3613..3630) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3619..3826 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3856..3996 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3856..3985 /label=AAV2 ITR rep_origin 4082..4537 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4169..4188) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4379..4400 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4604..4623) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4723..4745 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4783..4801) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4869..4973 /gene="bla" /label=AmpR promoter CDS 4974..5834 /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(5192..5211) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6005..6593 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6494..6513 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6747..6764 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtagatct gaattcggta cctagttatt aatagtaatc 181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc 541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt 601 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggcgcgcg ccaggcgggg 661 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga 721 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa 781 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg 841 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg 901 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt 961 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gctagagcct ctgctaacca 1021 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc 1081 tcatcatttt ggcaaagaat tcctcgaaga tccgaagggg ttcaagcttg ccaccatggt 1141 cgactcatca cgtcgtaagt ggaataagac aggtcacgca gtcagagcta taggtcggct 1201 gagctcactc gagaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcgaa 1261 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca 1321 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca 1381 gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 1441 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag 1501 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 1561 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta 1621 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 1681 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa 1741 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt 1801 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 1861 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 1921 caagctggag tacaacctgc cggaccaact gactgaagag cagatcgcag aatttaaaga 1981 ggaattctcc ctatttgaca aggacgggga tgggacaata acaaccaagg agctggggac 2041 ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca tgatcaatga 2101 agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa tgatggcaag 2161 aaaaatgaaa tacagggaca cggaagaaga aattagagaa gcgttcggtg tgtttgataa 2221 ggatggcaat ggctacatca gtgcagcaga gcttcgccac gtgatgacaa accttggaga 2281 gaagttaaca gatgaagagg ttgatgaaat gatcagggaa gcagacatcg atggggatgg 2341 tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aaggaattcg gaagcggagc 2401 tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg gacctgatat 2461 cgccaccatg ttgtttccaa gcaacactat cttgggggtt ttcgtcatat ctgagttttg 2521 tgtgggagtc ttagggaact cattgctcct catgctgtat atattcaact tctgggtcaa 2581 agctcgcttt aacagacaga tcgatcccat tttcatgcac ctgatgatag tcaatatgtt 2641 gacaatcata tttgctatga taccatatat cacatcatcc tttggagtgc ctagttttct 2701 gcatgatgct ggctgtaagg cctttgtgta tatctataga gtctcccggg ctatgtccat 2761 ctctagcact tctattctaa gcacattcca ggccatcacc atcactccca gtaattctaa 2821 gtgggcgtgg cttaagccta aactctctaa gtggactttt tgcaccttcc tgttttcctg 2881 gttcatcaac atggtcatat atgtgcacct gattgagaat gtaatagcaa aaactaatta 2941 cactggtgtt aattatggat attctcatgt ttactgtgaa agtagtccag ttgagtaccc 3001 caatccagga tttttcctcg gcgttatcat aacaagagac cttgtccttc tggtcatcat 3061 gatgtggacc agcctctcca tggtgaccct cctctacagg catcacaaga gagcccagca 3121 tctccgcaac ccaagactct cctctcagaa gtctccagaa cacaaagcca ctcacacaat 3181 cctgttgctt gtaaactgct ttgtgttctt ttatttgttg aacaacttaa tcacccttta 3241 tgataggttt tatacacgtg gaagaacagg aaatctgggg gcaattatca caattgtggc 3301 atcattttac cctactctct gcccattttt actaatgaac agtaataaaa tgtatttctt 3361 cccttcctat attcaaagag ccattggtga ctgatccaga catgataaga tacattgatg 3421 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 3481 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 3541 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttag tcgactagag 3601 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc 3661 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg 3721 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg 3781 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggagaga tctcacgtgc 3841 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc 3901 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg 3961 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcca gctggcgtaa 4021 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 4081 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 4141 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 4201 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 4261 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 4321 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 4381 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 4441 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 4501 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta 4561 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg 4621 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 4681 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 4741 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 4801 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 4861 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 4921 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 4981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 5041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 5101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 5161 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 5221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 5281 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 5341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 5401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 5461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 5521 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 5581 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 5641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 5701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 5761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 5821 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 5881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 5941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 6001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 6061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 6121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 6181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 6241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 6301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 6361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 6421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 6481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 6541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 6601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 6661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 6721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 6781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggggctcac 6841 atgt //