LOCUS       sequence-239604-        5097 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 5097)
  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 5097)
  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-fRGECO2
FEATURES             Location/Qualifiers
     source          1..5097
                     /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             904..921
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             925..957
                     /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(961..969,970..984)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1764..1783
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2274..2395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2285..2304)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2339..2358
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2576..3031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2663..2682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2873..2894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3098..3117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3217..3239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3277..3295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3363..3467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3468..3536,3537..4328)
                     /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(3686..3705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4499..5087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4988..5007
                     /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 ggaattcatg
      841 ctgcagaacg agcttgctct taagttggct ggacttgata ttaacaagac tggaggaggt
      901 tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg
      961 gatctgtacg acgatgacga taaggatctc gcaacaatgg tcgactcatc acgtcgtaag
     1021 tacaataagg caggtcacgc agtcagagct ataggtcggc tgagctcacc cgtggtttcc
     1081 gagcggatgt accccgagga cggcgccctg aagagcgaga tcaagaaggg gctgaggctg
     1141 aaggacggcg gccactacgc cgccgaggtc aagaccacct acaaggccaa gaagcccgtg
     1201 cagctgcccg gcgcctacat cgtggacatc aagttggaca tcgtgtccca caacgaggac
     1261 tacaccatcg tggaacagtg cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1321 gagctgtaca agggaggtac aggcgggagt ctggtgagca agggcgagga ggataacatg
     1381 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     1441 gagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca gaccgctaag
     1501 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     1561 atgtacggct ccaaggccta cattaagcac ccagccgaca tccccgacta cttcaagctg
     1621 tccttccccg agggcttcag gtgggagcgc gtgatgaact tcgaggacgg cggcattatt
     1681 cacgtgaacc aggactcctc cctgcaggac ggcgtattca tctacaaggt gaagctgcgc
     1741 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggct
     1801 acgcgtgacg acctgactga agagcagatc gcagaattta aagaggcttt ctccctattt
     1861 gacaaggacg gggatgggac gataacaacc aaggagcttg ggacggtgtt ccgatcgctg
     1921 gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt
     1981 gacggcacat tcgacttccc tgagttcctg acgatgatgg caagaaaaat gaatgacaca
     2041 gacagtgaag aggaaattcg cgaagcgttc cgcgtgtttg ataaggacgg caatggctac
     2101 atcggcgcag cagagcttcg ccacgtgatg acagaccttg gagagaagtt aacagatgag
     2161 gaggttgatg aaatgatcag ggtagcagcc atcgatgggg atggtcaggt aaactacgaa
     2221 gagtttgtac aaatgatgac agcgaagtag gcggccgctt cgagcagaca tgataagata
     2281 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2341 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2401 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     2461 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgggctggc gtaatagcga
     2521 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     2581 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2641 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2701 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2761 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2821 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2881 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2941 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3001 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3061 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3121 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3181 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3241 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3301 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3541 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3601 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3661 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3781 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3841 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3901 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4021 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4081 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4201 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4261 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4321 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4381 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4441 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4561 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4621 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4681 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4741 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4801 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4861 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4921 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4981 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5041 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagca
//
