LOCUS       sequence-264432-        5090 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 5090)
  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 5090)
  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..5090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..538
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          575..707
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     690..709
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        752..770
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             927..944
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             948..980
                     /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(984..992,993..1007)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1035..1091
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     polyA_signal    2270..2391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2335..2354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2572..3027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2659..2678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2869..2890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3094..3113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3213..3235
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3273..3291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3359..3463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3464..3532,3533..4324)
                     /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(3682..3701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4495..5083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4984..5003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac
       61 agatctaatc tgcagagggc cctgcgtatg agtgcaagtg ggttttaaga ccaggatgag
      121 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      181 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      241 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      301 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      361 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      421 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      481 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagct
      541 gcagctagga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
      601 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
      661 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
      721 gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg ctagcgctac
      781 cggactcaag cttggtaccg aactcggatc cactagtaac ggccgccagt gtgctggaat
      841 tcggcttggg gatatccacc atggtgcaga acgagcttgc tcttaagttg gctggacttg
      901 atattaacaa gactggagga ggttctcatc atcatcatca tcatggtatg gctagcatga
      961 ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat ctcgcaacaa
     1021 tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga gctataggtc
     1081 ggctgagctc agcgatcaac tgcgaaatga tgtacccagc ggatggtggt ctgcgtggtt
     1141 acactcacat ggcgctgaaa gttgatggcg gcggtcacct gtcctgttct ttcgtgacca
     1201 cctaccgctc caaaaagact gtcggcaaca ttaagatgcc tgccattcat agcgtcagcc
     1261 accgtctgga gcgcctggag gagagcgata acgaaatgtt tgtcgtacag cgtgaacacg
     1321 cagttgccaa gtttgtgggc ctgggtggtg gcggcggtac cggagggagc atgaactccc
     1381 tgatcaagga gaacatgcgt atgaaagtgg ttctggaagg ctccgtaaac ggccaccagt
     1441 tcaaatgcac tggtgaaggc gaaggcaacc cgtatatggg cacccagact atgcgtatca
     1501 aagtgatcga gggtggtccg ctgccgtttg cgttcgacat cctggcgacg tcctttatgt
     1561 atggctcccg taccttcatc aaatatccga aaggcatccc ggatttcttt aagcagtcct
     1621 tcccggaagg ttttacctgg gaacgtgtga cccgttacga agacggcggc gtaattaccg
     1681 ttatgcaaga cacgtctctg gaggatggct gcctggtgta tcacgtgcag gttcgcggtg
     1741 tgaacttccc gagcaatggt gctgtaatgc aaaagaaaac caaaggttgg gagcctacgg
     1801 actcccaaac cactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca
     1861 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc
     1921 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg
     1981 gcacaatcga cttccctgag ttcctgatta tgatggcaag aaaaatgaaa tacacagaca
     2041 gtgaagaaga aattagagaa gcgttcggcg tgtttgctaa ggatggcaat ggctacatca
     2101 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     2161 ttgatgaaat gatcagggaa gcagacagcg atggggatgg tcaggtaaac tacgaagagt
     2221 ttgtacaaat gatgacagcg aagtaggcgg ccgcttcgag cagacatgat aagatacatt
     2281 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2341 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2401 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     2461 taaaacctct acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa tagcgaagag
     2521 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     2581 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2641 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2701 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2761 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2821 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2881 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2941 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3001 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3061 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3121 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3181 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3241 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3421 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
//
