LOCUS Exported 4451 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of green fluorescent genetically encoded Ca2+ indicator for optical imaging (version 1.1). ACCESSION . VERSION . KEYWORDS CMV-G-GECO1.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4451) AUTHORS Zhao Y, Araki S, Wu J, Teramoto T, Chang YF, Nakano M, Abdelfattah AS, Fujiwara M, Ishihara T, Nagai T, Campbell RE TITLE An Expanded Palette of Genetically Encoded Ca2+ Indicators. JOURNAL Science. 2011 Sep 8. PUBMED 21903779 REFERENCE 2 (bases 1 to 4451) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4451 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 931..987 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 1200..1221 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1588..1607) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 1723..2166 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA ELQDMINEVDADGDGTIDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK" promoter complement(2227..2245) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2228..2245) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2265..2282) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2271..2495 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin complement(2696..3284) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2776..2795) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3455..4315) /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 4078..4097 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4316..4420) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 901 gagctcggat ccaccatggt cgactcatca cgtcgtaagt ggaataagac aggtcacgca 961 gtcagagcta taggtcggct gagctcactc gagaacgtct atatcaaggc cgacgagcag 1021 aagaacggca tcaaggcgta cttcaagatc cgccacaaca tcgagggcgg cggcgtgcag 1081 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 1141 aaccactacc tgagcgtgca gtccatactt tcgaaagacc ccaacgagaa gcgcgatcac 1201 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 1261 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg 1321 cccatccagg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 1381 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 1441 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 1501 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 1561 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 1621 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 1681 gacggcaaca tcctggggca caagctggag tacaacacgc gtgaccaact gactgaagag 1741 cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacgata 1801 acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag 1861 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag 1921 ttcctgacga tgatggcaag aaaaatgaat gacacagaca gtgaagagga aattagagaa 1981 gcgttccgtg tgtttgataa ggacggcaat ggctacatcg gcgcagcaga gcttcgccac 2041 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggta 2101 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg 2161 aagtaagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg 2221 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc 2281 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2341 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2401 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2461 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2521 gggctctagt ccggaactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 2581 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 2641 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 2701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 2761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 2821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 2881 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 2941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 3001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 3061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 3121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 3181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 3241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 3301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 3361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 3421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 3481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 3541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 3601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 3661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 3721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 3781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 3841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 3901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 3961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 4021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 4081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 4141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 4201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 4261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 4321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 4381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 4441 cacctgacgt c //