LOCUS       sequence-315961-        4454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carmine intensiometric genetically encoded Ca2+-indicators for 
            optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4454)
  AUTHORS   Wu J, Liu L, Matsuda T, Zhao Y, Rebane A, Drobizhev M, Chang YF, 
            Araki S, Arai Y, March K, Hughes TE, Sagou K, Miyata T, Nagai T, Li 
            WH, Campbell RE
  TITLE     Improved Orange and Red Ca Indicators and Photophysical 
            Considerations for Optogenetic Applications.
  JOURNAL   ACS Chem Neurosci. 2013 Mar 19.
  PUBMED    23452507
REFERENCE   2  (bases 1 to 4454)
  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: "ACS Chem 
            Neurosci. 2013 Mar 19."
COMMENT     Sequence Label: CMV-CAR-GECO1
FEATURES             Location/Qualifiers
     source          1..4454
                     /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"
     CDS             697..753
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1449..1468
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1492..1935
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     promoter        complement(1996..2014)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1997..2014)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2034..2051)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2040..2264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2465..3053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2545..2564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3224..4015,4016..4084))
                     /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     3847..3866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4085..4189)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4264..4283)
                     /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 tcggatccac catggtcgac tcatcacgtc gtaagtggaa taaggcaggt
      721 cacgcagtta gagctatagg tcggctgagc tcacccgtgg tttccgagcg gatgtacccc
      781 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac
      841 tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
      901 tacatcgtcg acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa
      961 cagtgcgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga
     1021 ggtacaggcg ggagtctggt gagcaagggc gtggaggata acatggccat cgtcaaggag
     1081 ttcatgcgct tcaagacgca catagagggc tccgtgaacg gccacgagtt cgagatcgag
     1141 ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag
     1201 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agattatgta cggctccaag
     1261 gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc
     1321 ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt taaccaggac
     1381 tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcccc
     1441 cccgacggcc ccgtaatgca gaagaagacc atgggctggg agcctacgcg tgaccaactg
     1501 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat
     1561 gggacgataa caaccaagga gctggggacg gtgatgcggt ctctggggca gaaccccaca
     1621 gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacattcgac
     1681 ttccctgagt tcctgacgat gatggcaaga aaaatgaatg acacagacag tgaagaggaa
     1741 attagagaag cgttccgcgt gtttgataag gacggcaatg gctacatcgg cgcagcagag
     1801 cttcgccacg tgatgacaga ccttggagag aagttaacag atgaggaggt tgatgaaatg
     1861 atcagggtag cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg
     1921 atgacagcga agtaggaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca
     1981 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     2041 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2101 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2161 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2221 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2281 aaccagctgg ggctctagtc cggaactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3121 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3181 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3241 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3301 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3361 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3421 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3481 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3541 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3601 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3661 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3721 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3781 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3841 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3901 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3961 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4021 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4081 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4141 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4201 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     4261 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     4321 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     4381 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     4441 cagatatacg cgtt
//
