LOCUS       sequence-264431-        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-fRGECO2
FEATURES             Location/Qualifiers
     source          1..5099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..542
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          579..711
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     694..713
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        756..774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             913..930
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             934..966
                     /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(970..978,979..993)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1773..1792
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2283..2404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2294..2313)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2348..2367
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2585..3040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2672..2691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2882..2903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3107..3126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3226..3248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3286..3304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3372..3476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3477..3545,3546..4337)
                     /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(3695..3714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4508..5096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4997..5016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
       61 cgacagatct aatctgcaga gggccctgcg tatgagtgca agtgggtttt aagaccagga
      121 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
      181 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
      241 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
      301 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
      361 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
      421 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
      481 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
      541 agctgcagct aggaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
      601 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
      661 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
      721 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcg
      781 ctaccggact caagcttggt accgaactcg gatccactag taacggccgc cagtgtgctg
      841 gaattcatgc tgcagaacga gcttgctctt aagttggctg gacttgatat taacaagact
      901 ggaggaggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa
      961 atgggtcggg atctgtacga cgatgacgat aaggatctcg caacaatggt cgactcatca
     1021 cgtcgtaagt acaataaggc aggtcacgca gtcagagcta taggtcggct gagctcaccc
     1081 gtggtttccg agcggatgta ccccgaggac ggcgccctga agagcgagat caagaagggg
     1141 ctgaggctga aggacggcgg ccactacgcc gccgaggtca agaccaccta caaggccaag
     1201 aagcccgtgc agctgcccgg cgcctacatc gtggacatca agttggacat cgtgtcccac
     1261 aacgaggact acaccatcgt ggaacagtgc gaacgcgccg agggccgcca ctccaccggc
     1321 ggcatggacg agctgtacaa gggaggtaca ggcgggagtc tggtgagcaa gggcgaggag
     1381 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1441 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga ggcctttcag
     1501 accgctaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1561 cctcagttca tgtacggctc caaggcctac attaagcacc cagccgacat ccccgactac
     1621 ttcaagctgt ccttccccga gggcttcagg tgggagcgcg tgatgaactt cgaggacggc
     1681 ggcattattc acgtgaacca ggactcctcc ctgcaggacg gcgtattcat ctacaaggtg
     1741 aagctgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     1801 tgggaggcta cgcgtgacga cctgactgaa gagcagatcg cagaatttaa agaggctttc
     1861 tccctatttg acaaggacgg ggatgggacg ataacaacca aggagcttgg gacggtgttc
     1921 cgatcgctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa tgaagtagat
     1981 gccgacggtg acggcacatt cgacttccct gagttcctga cgatgatggc aagaaaaatg
     2041 aatgacacag acagtgaaga ggaaattcgc gaagcgttcc gcgtgtttga taaggacggc
     2101 aatggctaca tcggcgcagc agagcttcgc cacgtgatga cagaccttgg agagaagtta
     2161 acagatgagg aggttgatga aatgatcagg gtagcagcca tcgatgggga tggtcaggta
     2221 aactacgaag agtttgtaca aatgatgaca gcgaagtagg cggccgcttc gagcagacat
     2281 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2341 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2401 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     2461 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgggctggcg
     2521 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2581 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2641 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2701 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2761 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2821 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2881 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2941 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3001 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     3061 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     3121 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3301 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3481 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3541 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3601 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3661 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3721 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3781 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3841 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3901 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3961 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4021 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4081 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4141 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4201 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4261 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4321 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4381 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4621 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaac
//
