LOCUS       sequence-264434-        5099 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 5099)
  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 5099)
  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-fRGECO1
FEATURES             Location/Qualifiers
     source          1..5099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        94..541
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          578..710
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     693..712
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        755..773
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             912..929
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             933..965
                     /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(969..977,978..992)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1020..1076
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1772..1791
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2282..2403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2293..2312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2347..2366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2584..3039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2671..2690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2881..2902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3106..3125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3225..3247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3285..3303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3371..3475
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3476..3544,3545..4336)
                     /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(3694..3713)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4507..5095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4996..5015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc
       61 gacagatcta atctgcagag ggccctgcgt atgagtgcaa gtgggtttta agaccaggat
      121 gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac
      181 ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg
      241 aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg
      301 cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca
      361 aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca
      421 ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga
      481 ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca
      541 gctgcagcta ggaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
      601 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
      661 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
      721 ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat aggctagcgc
      781 taccggactc aagcttggta ccgaactcgg atccactagt aacggccgcc agtgtgctgg
      841 aattcatgct gcagaacgag cttgctctta agttggctgg acttgatatt aacaagactg
      901 gaggaggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
      961 tgggtcggga tctgtacgac gatgacgata aggatctcgc aacaatggtc gactcatcac
     1021 gtcgtaagtg gaataaggca ggtcacgcag tcagagctat aggtcggctg agctcacccg
     1081 tggtttccga gcggatgtac cccgaggacg gcgccctgaa gagcgagatc aagaaggggc
     1141 tgaggctgaa ggacggcggc cactacgccg ccgaggtcaa gaccacctac aaggccaaga
     1201 agcccgtgca gctgcccggc gcctacatcg tggacatcaa gttggacatc gtgtcccaca
     1261 acgaggacta caccatcgtg gaacagtgcg aacgcgccga gggccgccac tccaccggcg
     1321 gcatggacga gctgtacaag ggaggtacag gcgggagtct ggtgagcaag ggcgaggagg
     1381 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1441 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag gcctttcaga
     1501 ccgctaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1561 ctcagttcat gtacggctcc aaggcctaca ttaagcaccc agccgacatc cccgactact
     1621 tcaagctgtc cttccccgag ggcttcaggt gggagcgcgt gatgaacttc gaggacggcg
     1681 gcattattca cgtgaaccag gactcctccc tgcaggacgg cgtattcatc tacaaggtga
     1741 agctgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct
     1801 gggaggctac gcgtgacgac ctgactgaag agcagatcgc agaatttaaa gaggctttct
     1861 ccctatttgc caaggacggg gatgggacga taacaaccaa ggagcttggg acggtgttcc
     1921 gatcgctggg gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg
     1981 ccgacggtga cggcacattc gacttccctg agttcctgac gatgatggca agaaaaatga
     2041 atgacacaga cagtgaagag gaaattcgcg aagcgttccg cgtgtttgat aaggacggca
     2101 atggctacat cggcgcagca gagcttcgcc acgtgatgac agaccttgga gagaagttaa
     2161 cagatgagga ggttgatgaa atgatcaggg tagcagacat cgatggggat ggtcaggtaa
     2221 actacgaaga gtttgtacaa atgatgacag cgaagtaggc ggccgcttcg agcagacatg
     2281 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2341 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2401 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2461 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gggctggcgt
     2521 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2581 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2641 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2701 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2761 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2821 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2881 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2941 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3001 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     3061 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     3121 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3181 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3241 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3301 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3361 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3421 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3481 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3541 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3601 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3661 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3721 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3781 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3841 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3901 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3961 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4021 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4081 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4141 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4201 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4261 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4321 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4381 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4441 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4501 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4561 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4621 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4681 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4741 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4801 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4861 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4921 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4981 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5041 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacg
//
