LOCUS       sequence-239609-        5088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses red fluorescent calcium sensor in hippocampal slices.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5088)
  AUTHORS   Kerruth S, Coates C, Durst CD, Oertner TG, Torok K
  TITLE     The kinetic mechanisms of fast-decay red-fluorescent genetically 
            encoded calcium indicators.
  JOURNAL   J Biol Chem. 2019 Mar 15;294(11):3934-3946. doi: 
            10.1074/jbc.RA118.004543. Epub 2019 Jan 16.
  PUBMED    30651353
REFERENCE   2  (bases 1 to 5088)
  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: 
            "10.1074/jbc.RA118.004543"; journalName: "J Biol Chem"; date: 
            "2019-03-15- 15"; volume: "294"; issue: "11"; pages: "3934-3946"
COMMENT     Sequence Label: pCI-syn-fRCaMP2
FEATURES             Location/Qualifiers
     source          1..5088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        86..533
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          570..702
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     685..704
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        747..765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             922..939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             943..975
                     /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             join(979..987,988..1002)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1030..1086
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     polyA_signal    2265..2386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2276..2295)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2330..2349
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2567..3022
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2654..2673)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2864..2885
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3089..3108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3208..3230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3268..3286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3354..3458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3459..3527,3528..4319)
                     /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(3677..3696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4490..5078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4979..4998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcggcctt ttacggttcc tggcctttgc tggccttttg ctcacatggc tcgacagatc
       61 taatctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg
      121 gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca acccccattc
      181 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt
      241 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc
      301 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc
      361 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga
      421 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc
      481 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagctgcagc
      541 taggaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
      601 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
      661 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
      721 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc gctaccggac
      781 tcaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc
      841 ttggggatat ccaccatggt gcagaacgag cttgctctta agttggctgg acttgatatt
      901 aacaagactg gaggaggttc tcatcatcat catcatcatg gtatggctag catgactggt
      961 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc aacaatggtc
     1021 gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg
     1081 agctcagcga tcaactgcga aatgatgtac ccagcggatg gtggtctgcg tggttacact
     1141 cacatggcgc tgaaagttga tggcggcggt cacctgtcct gttctttcgt gaccacctac
     1201 cgctccaaaa agactgtcgg caacattaag atgcctgcca ttcatagcgt cagccaccgt
     1261 ctggagcgcc tggaggagag cgataacgaa atgtttgtcg tacagcgtga acacgcagtt
     1321 gccaagtttg tgggcctggg tggtggcggc ggtaccggag ggagcatgaa ctccctgatc
     1381 aaggagaaca tgcgtatgaa agtggttctg gaaggctccg taaacggcca ccagttcaaa
     1441 tgcactggtg aaggcgaagg caacccgtat atgggcaccc agactatgcg tatcaaagtg
     1501 atcgagggtg gtccgctgcc gtttgcgttc gacatcctgg cgacgtcctt tatgtatggc
     1561 tcccgtacct tcatcaaata tccgaaaggc atcccggatt tctttaagca gtccttcccg
     1621 gaaggtttta cctgggaacg tgtgacccgt tacgaagacg gcggcgtaat taccgttatg
     1681 caagacacgt ctctggagga tggctgcctg gtgtatcacg tgcaggttcg cggtgtgaac
     1741 ttcccgagca atggtgctgt aatgcaaaag aaaaccaaag gttgggagcc tacggactcc
     1801 caaaccactg aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac
     1861 ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac
     1921 cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca
     1981 atcgacttcc ctgagttcct gattatgatg gcaagaaaaa tgaaatacac agacagtgaa
     2041 gaagaaatta gagaagcgtt cggcgtgttt gctaaggatg gcaatggcta catcagtgca
     2101 gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat
     2161 gaaatgatca gggaagcaga cagcgatggg gatggtcagg taaactacga agagtttgta
     2221 caaatgatga cagcgaagta ggcggccgct tcgagcagac atgataagat acattgatga
     2281 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2341 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2401 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     2461 cctctacaaa tgtggtaaaa tcgataagga tccgggctgg cgtaatagcg aagaggcccg
     2521 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     2581 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2641 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2701 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2761 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2821 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2881 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2941 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3001 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3061 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3121 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3181 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3241 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3301 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3421 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3481 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3541 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3601 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3661 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3721 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3781 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3841 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3901 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3961 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4021 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4081 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4141 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4201 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4261 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4321 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4381 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4621 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4681 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4741 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4801 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4861 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4921 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4981 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5041 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagca
//
