LOCUS       sequence-89442-e        4568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoded a yellow fluorescent calcium ion indicator with inverted 
            fluorescence response for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4568)
  AUTHORS   Zhao Y, Abdelfattah AS, Zhao Y, Ruangkittisakul A, Ballanyi K, 
            Campbell RE, Harrison DJ
  TITLE     Microfluidic cell sorter-aided directed evolution of a protein-based
            calcium ion indicator with an inverted fluorescent response.
  JOURNAL   Integr Biol (Camb). 2014 May 19.
  PUBMED    24840546
REFERENCE   2  (bases 1 to 4568)
  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"; journalName: "Integr Biol
            (Camb). 2014 May 19."
COMMENT     Sequence Label: CMV-Y-GECO1
FEATURES             Location/Qualifiers
     source          1..4568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             925..942
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             946..978
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1033..1089
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             2043..2057
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     promoter        complement(2344..2362)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2345..2362)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2382..2399)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2388..2612
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2893..2912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3572..4363,4364..4432))
                     /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     4195..4214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4433..4537)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ccaccatggg ttctcatcat catcatcatc atggtatggc tagcatgact
      961 ggtggacagc aaatgggtcg ggatctgtac gacgaagatg ataaggatct cgccacaatg
     1021 gccgactcat cacgtcgtaa gtggaataag acaggtcacg cagtcagagc tataggccgg
     1081 ctgagctcac tcgagggtta tagtaccgag aaaattgtgc ctgttccaaa gcaagggata
     1141 ttaaaaggaa aggtcagcat gtacctattg ctgaaggatg gcggtcgtta tcattgccag
     1201 tttgatacgg tatataaagc taaatcagtg ccctccaaga tgccaggacg gcattttata
     1261 cagcataagc tccttaggga ggaccgctct gatgctaaaa accataaatg gcaattaaca
     1321 gaacatgcaa tcgcaggcat ggacgagctg tacaagggag gtacaggcgg gagtatggtg
     1381 agcaagggcg aggggcaatc gaagcatggg ctaaaggagg agatgacggt gaagtaccac
     1441 atggaaggtt gcgttaatgg tcacaaattt gtcattactg gagagggcat tggaaaccct
     1501 tttaagggta agcaaactgc aaatttgtgt gtgatagaag gaggcccgct gccgttctcg
     1561 gaggacattt taagtccggg ttttaaatat ggtgaccgga ttttcacaga gtacccacag
     1621 gatattgtag attacttcaa gaactcatgt ccagagggct atacgtggga aaggagcttc
     1681 ttctatgagg acggagcggt ctgtcgatgc aacgcggaca taacagtctc tgaaaaggag
     1741 aactgcatct atcacaaaag tatcttctat ggggtgaatt ttcccgccga cggccccgta
     1801 atgaagaaaa tgaccactaa ttgggaagct gacacgcgtg accaactgac tgaagagcag
     1861 atcgcagaat ttaaagaggc tttctcccta tttgacaagg acggggatgg gacgatgaca
     1921 accgaggagc tggggacggt gctgcggtct ctggggcaga acccctcaga agcagagctg
     1981 cgggacatga tcaatgaagt agatgccgac ggtgacggca caatcggctt ccctgagttc
     2041 ctgacgatga tgacaagaaa aatgaatgac acggacagtg aagaggaaat tagagaagcg
     2101 ttccgtgtgt ttgataagga cggcaatggc tacatcggcg cagcagagct tcgccacgtg
     2161 atgacaaacc ttggagagaa gttaacagat gaagagtttg atgaaatgat cagggtagca
     2221 gactccgacg gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag
     2281 taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     2341 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     2401 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2461 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2521 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2581 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2641 ctctagtccg gaactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2701 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3001 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3241 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4561 ctgacgtc
//
