LOCUS Exported 5455 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Improved genetic encoded calcium sensor based on GCaMP6m. Also called GCaMP-X. ACCESSION . VERSION . KEYWORDS pN1-GCaMP6m-XC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5455) AUTHORS Yang Y, Liu N, He Y, Liu Y, Ge L, Zou L, Song S, Xiong W, Liu X TITLE Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP. JOURNAL Nat Commun. 2018 Apr 17;9(1):1504. doi: 10.1038/s41467-018-03719-6. PUBMED 29666364 REFERENCE 2 (bases 1 to 5455) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5455 /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 616..625 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 733..2082 /codon_start=1 /product="improved fluorescent protein-based calcium sensor (Chen et al., 2013)" /label=GCaMP6m /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" regulatory 829..838 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 1119..1140 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1246..1267) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1507..1526) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" polyA_signal 2243..2364 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2280..2299) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2334..2353 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2371..2826) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2508..2529) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2720..2739 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2853..2957 /gene="bla" /label=AmpR promoter promoter 2959..3316 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2982..3002) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3167..3302 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3229..3248 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3351..4145 /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(3405..3424) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4015..4034 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4333..4352) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4377..4424 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4753..5341 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5242..5261 /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 ccggactcag atctcgccac catgggcatg gacgagctgt acaagggtac cgctgcaacc 661 aaaattcagg ctgccttccg tggacacata acaaggaaaa agctcaaaga cgagaagaag 721 ggtgctgaat tcatgggttc tcatcatcat catcatcatg gtatggctag catgactggt 781 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc 841 gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg 901 agctcactcg agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac 961 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag 1021 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag 1081 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 1141 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc 1201 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 1261 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac 1321 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 1381 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 1441 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc 1501 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 1561 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 1621 aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga atttaaagag 1681 gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg 1741 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa 1801 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga 1861 aaagggagct acagggacac ggaagaagaa attagagaag cgttcggtgt gtttgataag 1921 gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag 1981 aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt 2041 caggtaaact acgaagagtt tgtacaaatg atgacagcga agcttgctct taagttggct 2101 ggacttgata ttggttctta agcggccgcg actctagatc ataatcagcc ataccacatt 2161 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa 2221 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag 2281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 2341 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat 2401 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 2461 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 2521 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 2581 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 2641 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 2701 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 2761 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 2821 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 2881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 2941 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg 3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 3061 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 3121 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 3181 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 3241 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 3301 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac 3361 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3421 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3481 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg 3541 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 3601 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 3661 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 3721 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 3781 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 3841 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc 3901 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 3961 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 4021 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 4081 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 4141 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 4201 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4261 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag 4321 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc 4381 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg 4441 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 4501 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 4561 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga 4621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 4681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 4741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 4801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 4861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 4921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 4981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 5041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 5101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 5161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 5221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 5281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 5341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 5401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat //