LOCUS       sequence-264454-        5361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses red fluorescent calcium sensor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5361)
  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 5361)
  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: pGP-CMV-NES-fRGECO1
FEATURES             Location/Qualifiers
     source          1..5361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             692..709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             713..745
                     /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(749..757,758..772)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             800..856
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1552..1571
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2160..2281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2197..2216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2251..2270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2288..2743)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2425..2446)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2637..2656
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2770..2874
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2876..3233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2876..3067
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2899..2919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3084..3219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3268..4062
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3268..4062
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3322..3341)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3932..3951
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4250..4269)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4294..4341
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4670..5258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5159..5178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctatgct gcagaacgag cttgctctta agttggctgg
      661 acttgatatt aacaagactg gaggaggttc tcatcatcat catcatcatg gtatggctag
      721 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc
      781 aacaatggtc gactcatcac gtcgtaagtg gaataaggca ggtcacgcag tcagagctat
      841 aggtcggctg agctcacccg tggtttccga gcggatgtac cccgaggacg gcgccctgaa
      901 gagcgagatc aagaaggggc tgaggctgaa ggacggcggc cactacgccg ccgaggtcaa
      961 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacatcg tggacatcaa
     1021 gttggacatc gtgtcccaca acgaggacta caccatcgtg gaacagtgcg aacgcgccga
     1081 gggccgccac tccaccggcg gcatggacga gctgtacaag ggaggtacag gcgggagtct
     1141 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1201 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1261 cccctacgag gcctttcaga ccgctaagct gaaggtgacc aagggtggcc ccctgccctt
     1321 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctaca ttaagcaccc
     1381 agccgacatc cccgactact tcaagctgtc cttccccgag ggcttcaggt gggagcgcgt
     1441 gatgaacttc gaggacggcg gcattattca cgtgaaccag gactcctccc tgcaggacgg
     1501 cgtattcatc tacaaggtga agctgcgcgg caccaacttc ccccccgacg gccccgtaat
     1561 gcagaagaag accatgggct gggaggctac gcgtgacgac ctgactgaag agcagatcgc
     1621 agaatttaaa gaggctttct ccctatttgc caaggacggg gatgggacga taacaaccaa
     1681 ggagcttggg acggtgttcc gatcgctggg gcagaacccc acagaagcag agctgcagga
     1741 catgatcaat gaagtagatg ccgacggtga cggcacattc gacttccctg agttcctgac
     1801 gatgatggca agaaaaatga atgacacaga cagtgaagag gaaattcgcg aagcgttccg
     1861 cgtgtttgat aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac
     1921 agaccttgga gagaagttaa cagatgagga ggttgatgaa atgatcaggg tagcagacat
     1981 cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtaggc
     2041 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2101 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2281 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2341 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2401 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2461 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2521 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2581 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2641 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2701 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     2761 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2821 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     2881 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     3241 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3301 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3361 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3421 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     3481 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3541 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3601 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3661 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3721 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3781 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3841 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3901 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3961 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4021 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4081 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4141 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4201 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     4261 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4321 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4381 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4441 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4501 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4561 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4621 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4681 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4741 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4801 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4861 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4921 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4981 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5041 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5101 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5161 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5221 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5281 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5341 attctgtgga taaccgtatt a
//
