LOCUS       sequence-122742-        4451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of green fluorescent genetically encoded Ca2+ 
            indicator for optical imaging (version 1.0).
ACCESSION   .
VERSION     .
KEYWORDS    .
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 30;333(6051):1888-91. doi: 10.1126/science.1208592
  PUBMED    21903779
REFERENCE   2  (bases 1 to 4451)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2011-09-30- 30"; volume: "333"; 
            issue: "6051"; pages: "1888-91"
COMMENT     Sequence Label: CMV-G-GECO1.0
FEATURES             Location/Qualifiers
     source          1..4451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    42..266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     255..272
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        292..310
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     292..309
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(371..814)
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     primer_bind     930..949
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1316..1337)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(1550..1606)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(1655..1674)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1656..1674)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1719..1922)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1748..1768)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1923..2302
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2474..2493
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2568..2672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2673..2741,2742..3533)
                     /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(2891..2910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3704..4292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4193..4212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttccggacta gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
       61 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
      121 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
      181 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
      241 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctag catttaggtg
      301 acactataga atagggccct ctagatgcat gctcgagcgg ccgccagtgt gatggatatc
      361 tgcagaattc ttacttcgct gtcatcattt gtacaaactc ttcgtagttt acctgaccat
      421 ccccatcgat gtctgctacc ctgatcattt catcaacctc ttcatctgtt aacttctctc
      481 caaggtttgt catcacgtgg cgaagctctg ctgcgccgat gtagccattg ccgtccttat
      541 caaacacacg gaacgcttct ctaatttctt cttcactgtc tgtgtctttc atttttcttg
      601 ccatcattgt caggaactca gggaagtcga ttgtgccgtc accgtcggca tctacttcat
      661 tgatcatgtc ctgcagctct gcttctgtgg ggttctgccc cagagaccgc atcaccgtcc
      721 ccagctcctt ggttgttatc gtcccatccc cgtccttgtc aaatagggag aaagcctctt
      781 taaattctgc gatctgctct tcagtcagtt ggtcacgcgt gttgtactcc agcttgtgcc
      841 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
      901 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
      961 tggtgcgctc ctggatgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     1021 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     1081 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     1141 tggcatcgcc ctcaccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     1201 gctcgacctg gatgggcacc accccggtga acagctcctc gcccttgctc accatgctcc
     1261 ctccggtacc gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     1321 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttcgaaagt atggactgca
     1381 cgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     1441 ggtagtggta ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttcg
     1501 ccttgatgcc gttcttctgc ttgtcggcct tgatatagac gttctcgagt gagctcagcc
     1561 gacctatagc tctgactgcg tgacctgtct tattccactt acgacgtgat gagtcgacca
     1621 tggtggatcc gagctcggta ccaagcttgg gtctccctat agtgagtcgt attaatttcg
     1681 ataagccagt aagcagtggg ttctctagtt agccagagag ctctgcttat atagacctcc
     1741 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     1801 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     1861 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     1921 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     1981 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2041 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     2101 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     2161 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     2221 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     2281 taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca
     2341 aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta
     2401 gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac
     2461 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag gggatcggga
     2521 gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2581 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2641 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2701 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2761 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2821 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2881 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2941 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3001 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3061 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3121 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3181 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3241 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3301 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3361 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3421 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3481 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3541 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3601 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3661 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3721 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3781 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3841 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3901 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3961 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4021 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4081 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4141 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4201 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4261 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4321 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4381 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4441 agcgcagcga g
//
