LOCUS       sequence-239614-        5341 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 5341)
  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 5341)
  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-fRCaMP2
FEATURES             Location/Qualifiers
     source          1..5341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             688..705
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             709..741
                     /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(745..753,754..768)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             796..852
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     polyA_signal    2129..2250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2166..2185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2220..2239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2257..2712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2394..2415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2606..2625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2739..2843
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2845..3202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2845..3036
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2868..2888)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3053..3188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3237..4031
                     /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             3237..4031
                     /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(3291..3310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3901..3920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4219..4238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4263..4310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4639..5227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5128..5147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgccac catgctgcag aacgagcttg ctcttaagtt ggctggactt
      661 gatattaaca agactggagg aggttctcat catcatcatc atcatggtat ggctagcatg
      721 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgcaaca
      781 atggtcgact catcacgtcg taagtggaat aagacaggtc acgcagtcag agctataggt
      841 cggctgagct cagcgatcaa ctgcgaaatg atgtacccag cggatggtgg tctgcgtggt
      901 tacactcaca tggcgctgaa agttgatggc ggcggtcacc tgtcctgttc tttcgtgacc
      961 acctaccgct ccaaaaagac tgtcggcaac attaagatgc ctgccattca tagcgtcagc
     1021 caccgtctgg agcgcctgga ggagagcgat aacgaaatgt ttgtcgtaca gcgtgaacac
     1081 gcagttgcca agtttgtggg cctgggtggt ggcggcggta ccggagggag catgaactcc
     1141 ctgatcaagg agaacatgcg tatgaaagtg gttctggaag gctccgtaaa cggccaccag
     1201 ttcaaatgca ctggtgaagg cgaaggcaac ccgtatatgg gcacccagac tatgcgtatc
     1261 aaagtgatcg agggtggtcc gctgccgttt gcgttcgaca tcctggcgac gtcctttatg
     1321 tatggctccc gtaccttcat caaatatccg aaaggcatcc cggatttctt taagcagtcc
     1381 ttcccggaag gttttacctg ggaacgtgtg acccgttacg aagacggcgg cgtaattacc
     1441 gttatgcaag acacgtctct ggaggatggc tgcctggtgt atcacgtgca ggttcgcggt
     1501 gtgaacttcc cgagcaatgg tgctgtaatg caaaagaaaa ccaaaggttg ggagcctacg
     1561 gactcccaaa ccactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac
     1621 aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg
     1681 cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac
     1741 ggcacaatcg acttccctga gttcctgatt atgatggcaa gaaaaatgaa atacacagac
     1801 agtgaagaag aaattagaga agcgttcggc gtgtttgcta aggatggcaa tggctacatc
     1861 agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag
     1921 gttgatgaaa tgatcaggga agcagacagc gatggggatg gtcaggtaaa ctacgaagag
     1981 tttgtacaaa tgatgacagc gaagtaggcg gccgcgactc tagatcataa tcagccatac
     2041 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2101 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2161 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2221 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     2281 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2341 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2401 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2461 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2521 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2581 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2641 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2701 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2761 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2821 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2881 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2941 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3181 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3241 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3301 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3361 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3421 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3481 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3541 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3601 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3661 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3721 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3781 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3841 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3901 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3961 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4021 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4081 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4141 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4201 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4261 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4321 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4381 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4441 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     4501 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4741 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5341 t
//
