LOCUS Exported 5046 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Soluble red fluorescent activator of Orai1 channels. ACCESSION . VERSION . KEYWORDS mCherry-CAD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5046) AUTHORS Dynes JL, Amcheslavsky A, Cahalan MD TITLE Genetically targeted single-channel optical recording reveals multiple Orai1 gating states and oscillations in calcium influx. JOURNAL Proc Natl Acad Sci U S A. 2016 Jan 12;113(2):440-5. doi: 10.1073/pnas.1523410113. Epub 2015 Dec 28. PUBMED 26712003 REFERENCE 2 (bases 1 to 5046) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5046 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1320 /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(758..776) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1026..1045 /label=mCherry-F /note="mCherry, forward primer" CDS 1684..1701 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 1834..1955 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1871..1890) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1925..1944 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1962..2417) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2099..2120) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2311..2330 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2444..2548 /gene="bla" /label=AmpR promoter promoter 2550..2907 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2573..2593) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2758..2893 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2820..2839 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2942..3736 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2996..3015) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3606..3625 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3924..3943) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3968..4015 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4344..4932 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4833..4852 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc 661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt 781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc 841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc 901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc 1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac 1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc 1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc 1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg 1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag 1321 tccggactca gatctcgagc tcaagcttcg aattcttatg ctccagaggc ccttcagaag 1381 tggctgcagc tgacacatga ggtggaggtg caatattaca acatcaagaa gcaaaatgct 1441 gagaagcagc tgctggtggc caaggagggg gctgagaaga taaaaaagaa gagaaacaca 1501 ctctttggca ccttccacgt ggcccacagc tcttccctgg atgatgtaga tcataaaatt 1561 ctaacagcta agcaagcact gagcgaggtg acagcagcat tgcgggagcg cctgcaccgc 1621 tggcaacaga tcgagatcct ctgtggcttc cagattgtca acaaccctgg catccacgga 1681 ggtcatcatc accatcacca ttgaggatcc accggatcta gataactgat cataatcagc 1741 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 1801 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 1861 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 1921 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat 1981 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 2041 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 2101 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 2161 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 2221 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 2281 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 2341 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 2401 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 2461 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 2521 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg 2581 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2641 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 2701 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 2761 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 2821 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 2881 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg 2941 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 3001 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 3061 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 3121 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 3181 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 3241 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 3301 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 3361 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 3421 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga 3481 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 3541 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 3601 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 3661 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 3721 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac 3781 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 3841 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 3901 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc 3961 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa 4021 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 4081 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 4141 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata 4201 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 4261 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 4321 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 4381 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 4441 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 4501 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 4561 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 4621 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 4681 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 4741 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 4801 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 4861 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 4921 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 4981 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 5041 atgcat //