LOCUS       sequence-227508-        4100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive bacterial expression of red fluorescent Ca2+ sensor 
            jRGECO1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4100)
  AUTHORS   Molina RS, Qian Y, Wu J, Shen Y, Campbell RE, Drobizhev M, Hughes TE
  TITLE     Understanding the Fluorescence Change in Red Genetically Encoded 
            Calcium Ion Indicators.
  JOURNAL   Biophys J. 2019 May 21;116(10):1873-1886. doi: 
            10.1016/j.bpj.2019.04.007. Epub 2019 Apr 13.
  PUBMED    31054773
REFERENCE   2  (bases 1 to 4100)
  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: 
            "10.1016/j.bpj.2019.04.007"; journalName: "Biophys J"; date: 
            "2019-05-21- 21"; volume: "116"; issue: "10"; pages: "1873-1886"
COMMENT     Sequence Label: pRSM42
FEATURES             Location/Qualifiers
     source          1..4100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4..51
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      148..603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(235..254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     445..466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        629..733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             734..1528
                     /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             734..1528
                     /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(788..807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1398..1417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      1699..2287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2188..2207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2441..2458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2543..2562
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2543..2561
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2645..2667
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2687..2704
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2708..2740
                     /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(2744..2752,2753..2767)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2789..2845
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     3541..3560
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(join(4090..4100,1..8))
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
       61 ggaactatat ccggatctgg cgtaatagcg aagaggccag caccgatcgc ccttcccaac
      121 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
      181 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      241 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      301 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      361 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      421 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
      481 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      541 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
      601 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      661 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      721 aaaaaggaag agtatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      781 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      841 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      901 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      961 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     1021 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     1081 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     1141 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     1201 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     1261 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     1321 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1381 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     1441 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     1501 ctatcgcctt cttgacgagt tcttctgact gtcagaccaa gtttactcat atatacttta
     1561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1681 aaggatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2101 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2461 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2521 caggatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
     2581 tctagaatca ccgagtttat tcttgacacc tgatgcgatg aatgatataa taggaaagta
     2641 ctgttttgtt taactttaag aaggagatat acatatgcgg ggttctcatc atcatcatca
     2701 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
     2761 cgataaggat ccgatggtcg actcatcacg tcgtaagtgg aataaggcag gtcacgcagt
     2821 cagagctata ggtcggctga gctcacccgt ggtttccgag cggatgtacc ccgaggacgg
     2881 cgccctgaag agcgagatca agaaggggct gaggctgaag gacggcggcc actacgccgc
     2941 cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt
     3001 ggacatcaag ttggacatcg tgtcccacaa cgaggactac accatcgtgg aacagtgcga
     3061 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg gaggtacagg
     3121 cgggagtctg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     3181 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     3241 cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca agggtggccc
     3301 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacat
     3361 taagcaccca gccgacatcc ccgactactt caagctgtcc ttccccgagg gcttcaggtg
     3421 ggagcgcgtg atgaacttcg aggacggcgg cattattcac gtgaaccagg actcctccct
     3481 gcaggacggc gtattcatct acaaggtgaa gctgcgcggc accaacttcc cccccgacgg
     3541 ccccgtaatg cagaagaaga ccatgggctg ggaggctacg cgtgacgacc tgactgaaga
     3601 gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg atgggacgat
     3661 aacaaccaag gagcttggga cggtgttccg atcgctgggg cagaacccca cagaagcaga
     3721 gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacattcg acttccctga
     3781 gttcctgacg atgatggcaa gaaaaatgaa tgacacagac agtgaagagg aaattcgcga
     3841 agcgttccgc gtgtttgata aggacggcaa tggctacatc ggcgcagcag agcttcgcca
     3901 cgtgatgaca gaccttggag agaagttaac agatgaggag gttgatgaaa tgatcagggt
     3961 agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc
     4021 gaagtaggaa ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4081 ctgctgccac cgctgagcaa
//
