pGP-CMV-Gcamp6f

LOCUS pGP-CMV-GCaMP6f 5308 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION Mammalian expression of ultrasensitive protein calcium sensor. ACCESSION . VERSION . KEYWORDS pGP-CMV-GCaMP6f SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5308) AUTHORS Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, Kim DS TITLE Ultrasensitive fluorescent proteins for imaging neuronal activity. JOURNAL Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. PUBMED 23868258 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pGP-CMV-Gcamp6f| FEATURES Location/Qualifiers source 1..5308 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(39..58) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 285..305 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(330..434) /gene="bla" /label=AmpR promoter rep_origin 461..916 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(548..567) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 758..779 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 923..1044 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(934..953) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 988..1007 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(1166..2518) /codon_start=1 /product="improved fluorescent protein-based calcium sensor (Chen et al., 2013)" /label=GCaMP6f /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" primer_bind 1725..1744 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1984..2005 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2111..2132) /label=EGFP-C /note="EGFP, forward primer" regulatory 2413..2422 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2515..2524 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(2572..2775) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(2601..2621) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2776..3079 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(3254..3842) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3334..3353) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 4171..4218 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 4243..4262 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(4450..5244) /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(4561..4580) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 5171..5190 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(5279..328) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5293..120 /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa 61 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat 121 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact 181 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat 241 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac 301 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt 361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca 481 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 541 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 601 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 661 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 721 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 781 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 841 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 901 ttaacgctta caatttacgc cttaagatac attgatgagt ttggacaaac cacaactaga 961 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc 1021 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt 1081 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct 1141 gattatgatc tagagtcgcg gccgctcact tcgctgtcat catttgtaca aactcttcgt 1201 agtttacctg accatcccca tcgatgtctg cttccctgat catttcatca acctcttcat 1261 ctgttaactt ctctccaagg tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc 1321 cattgccatc cttatcaaac acaccgaacg cttctctaat ttcttcttcc gtgtccctgt 1381 atttcatttt tcttgccatc attgtcagga actcagggaa gtcgattgtg ccgtcaccgt 1441 cggcatctac ttcattgatc atgtcctgca gctctgcttc tgtggggttc tgccccagag 1501 accgcatcac cgtccccagc tccttggttg ttattgtccc atccccgtcc ttgtcaaata 1561 gggagaattc ctctttaaat tctgcgatct gctcttcagt cagttggtcc ggcaggttgt 1621 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga 1681 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt 1741 cgtccttgaa gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga 1801 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg 1861 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg 1921 tcagcttgcc gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt 1981 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct 2041 tgctcaccat gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc 2101 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg 2161 aaagtttgga ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga 2221 tgggggtgtt ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc 2281 ggatcttgaa gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgttct 2341 cgagtgagct cagccgacct atagctctga ctgcgtgacc tgtcttattc cacttacgac 2401 gtgatgagtc gaccatggtg gcgagatcct tatcgtcatc gtcgtacaga tcccgaccca 2461 tttgctgtcc accagtcatg ctagccatac catgatgatg atgatgatga gaacccatgg 2521 tggcgagatc tgagtccggt agcgctagcg gatctgacgg ttcactaaac cagctctgct 2581 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt 2641 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat 2701 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg 2761 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg 2821 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac 2881 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta 2941 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac 3001 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt 3061 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat 3121 tgattactat taataactaa tgcatggcgg taatacggtt atccacagaa tcaggggata 3181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 3241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 3301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 3361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 3421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 3481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 3541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 3601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 3661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 3721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 3961 aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg 4021 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg 4081 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt 4141 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat 4201 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt 4261 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga 4321 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg 4381 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca 4441 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg 4501 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc 4561 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca 4621 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc 4681 atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc 4741 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc 4801 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc 4861 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg 4921 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct 4981 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca 5041 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac 5101 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat 5161 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc 5221 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt 5281 tgcaaaagcc taggcctcca aaaaagcc //