LOCUS Exported 6039 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Improved genetic encoded calcium sensor based on GCaMP6m. ACCESSION . VERSION . KEYWORDS AAV-syn-GCaMP6m-XC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6039) 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 6039) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6039 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 132..579 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 599..608 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 716..2065 /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 812..821 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 1102..1123 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1229..1250) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1490..1509) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 2123..2711 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2176..2196) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2594..2605) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2743..3219 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2942..2961) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3259..3399 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3259..3388 /label=AAV2 ITR rep_origin 3474..3929 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3561..3580) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3771..3792 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3946..3965) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4065..4087 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4125..4143) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4211..4315 /gene="bla" /label=AmpR promoter CDS 4316..5176 /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(4534..4553) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5347..5935 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5836..5855 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 5997..87 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag 61 tggccaactc catcactagg ggttcctgcg gccgcacgcg tgtgtctaga ctgcagaggg 121 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac 181 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat 241 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct 301 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc 361 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt 421 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg 481 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg 541 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaaggta ccggatccgc 601 caccatgggc atggacgagc tgtacaaggg taccgctgca accaaaattc aggctgcctt 661 ccgtggacac ataacaagga aaaagctcaa agacgagaag aagggtgctg aattcatggg 721 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg 781 ggatctgtac gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa 841 gtggaataag acaggtcacg cagtcagagc tataggtcgg ctgagctcac tcgagaacgt 901 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa 961 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga 1021 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga 1081 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac 1141 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga 1201 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca 1261 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa 1321 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac 1381 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa 1441 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa 1501 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct 1561 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct 1621 gccggaccaa ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga 1681 caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg 1741 gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga 1801 cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaggga gctacaggga 1861 cacggaagaa gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat 1921 cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga 1981 ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga 2041 gtttgtacaa atgatgacag cgaagcttgc tcttaagttg gctggacttg atattggttc 2101 ttaagatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac 2161 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 2221 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 2281 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 2341 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 2401 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 2461 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 2521 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt 2581 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 2641 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 2701 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac 2761 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc 2821 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg 2881 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg 2941 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc 3001 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc 3061 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc 3121 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg 3181 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg 3241 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg 3301 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg 3361 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc 3421 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc 3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 3541 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 3661 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc 3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 3781 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat 3841 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 3901 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga 3961 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc 4021 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg 4081 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct 4141 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg 4201 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 4261 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 4321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 4381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 4441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 4501 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 4561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 4621 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 4681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 4741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 4801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 4861 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 4921 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 4981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 5041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 5101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 5161 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg 6001 caggcagctg cgcgctcgct cgctcactga ggccgcccg //