LOCUS       sequence-292724-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Ca2+ inside primary cilia.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  AUTHORS   Su S, Phua SC, Derose R, Chiba S, Narita K, Kalugin PN, Katada T, 
            Kontani K, Takeda S, Inoue T
  TITLE     Genetically encoded calcium indicator illuminates calcium dynamics 
            in primary cilia.
  JOURNAL   Nat Methods. 2013 Sep 22. doi: 10.1038/nmeth.2647.
  PUBMED    24056873
REFERENCE   2  (bases 1 to 5807)
  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: "Nat 
            Methods. 2013 Sep 22. doi: 10.1038/nmeth.2647."
COMMENT     Sequence Label: 5HT6-G-GECO1.0
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2053..2109
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     2322..2343
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2710..2729)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2845..3288
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     promoter        complement(3349..3367)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3350..3367)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3387..3404)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3393..3617
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3818..4406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3898..3917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4577..5368,5369..5437))
                     /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     5200..5219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5438..5542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5617..5636)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccgc caccatggtt ccagagcccg gccctgtcaa cagtagcacc
      721 ccagcctggg gccccgggcc accgccagcc ccagggggca gcggctgggt ggccgctgcg
      781 ctgtgcgtgg tcatcgtgct gacggcagcc gccaattcgc tgctgatcgc gctcatctgc
      841 acgcagcccg cgctgcgcaa cacgtctaac ttcttcctgg tgtcgctctt cacgtcggac
      901 ctgatggtgg gattggtggt gatgccccca gccatgctga acgcgctgta tgggcgctgg
      961 gtactagctc gcggcctctg tctgctttgg accgccttcg acgtgatgtg ctgcagcgcc
     1021 tccattctca acctctgcct catcagcctg gaccgctacc tgctcatcct ctcgccgctg
     1081 cgctacaagc tgcgcatgac agccccgcgt gccctggcgc tcatcctggg tgcctggagc
     1141 ctcgccgcgc tggcctcctt cctgcccctc ttgctgggct ggcatgaact gggcaaagct
     1201 cgaacatctg ccccgggcca gtgccgccta ttggccagcc tgccttatgt cctcgtggcg
     1261 tccggcgtca cctttttcct gccttcgggt gccatctgct tcacctactg caggatcttg
     1321 ctggctgccc gcaagcaggc ggtgcaagtg gcctcgctca ccacgggcac ggctacggct
     1381 ggccaggcct tggaaacctt gcaggtgccc aggacaccac gcccagggat ggagtctgct
     1441 gacagtaggc gtctgaccac caagcatagc aggaaggcct tgaaggccag cctgaccctc
     1501 ggcatcctgc tgagcatgtt ctttgtcacc tggctgccct tctttgtggc cagcatagct
     1561 caggccgtat gtgactgcat ctctccaggt ctcttcgatg tcctcacatg gctgggatac
     1621 tgtaatagca ccatgaaccc tatcatctat cccctcttca tgcgggactt caagagggcc
     1681 ctgggcaggt tcgtgccgtg tgtccactgt cccccggagc accgggccag ccccgcctcc
     1741 ccctccatgt ggacctctca cagtggtgcc aggccaggcc tcagcctgca gcaggtgctg
     1801 cccctgcctc tgccacccaa ctcagattca gactcagctt cagggggcac ctcgggcctg
     1861 cagctcacag cccagctttt gctgcctgga gaggcgaccc gggacccccc gccacccacc
     1921 agggccccta ctgtggtcaa cttcttcgtc acagactctg tggagcctga gatacggcag
     1981 catccacttg gttcccccat gaacagtgct ggtggtagcg caggaggagg atccaccatg
     2041 gtcgactcat cacgtcgtaa gtggaataag acaggtcacg cagtcagagc tataggtcgg
     2101 ctgagctcac tcgagaacgt ctatatcaag gccgacaagc agaagaacgg catcaaggcg
     2161 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccta ccactaccag
     2221 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcgtg
     2281 cagtccatac tttcgaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2341 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg taccggaggg
     2401 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcca ggtcgagctg
     2461 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg cgatgccacc
     2521 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2581 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2641 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acatccagga gcgcaccatc
     2701 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2761 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2821 cacaagctgg agtacaacac gcgtgaccaa ctgactgaag agcagatcgc agaatttaaa
     2881 gaggctttct ccctatttga caaggacggg gatgggacga taacaaccaa ggagctgggg
     2941 acggtgatgc ggtctctggg gcagaacccc acagaagcag agctgcagga catgatcaat
     3001 gaagtagatg ccgacggtga cggcacaatc gacttccctg agttcctgac aatgatggca
     3061 agaaaaatga aagacacaga cagtgaagaa gaaattagag aagcgttccg tgtgtttgat
     3121 aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac aaaccttgga
     3181 gagaagttaa cagatgaaga ggttgatgaa atgatcaggg tagcagacat cgatggggat
     3241 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtaaga attctgcaga
     3301 tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca
     3361 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3421 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3481 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3541 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3601 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gtccggaact
     3661 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4501 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4561 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4621 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4681 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4741 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4801 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4861 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4921 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4981 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5041 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5101 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5161 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5221 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5281 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5341 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5401 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5581 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     5641 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     5701 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     5761 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
