LOCUS       sequence-220795-        4100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive bacterial expression of red fluorescent Ca2+ sensor 
            REX-GECO1.
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: pRSM41
FEATURES             Location/Qualifiers
     source          1..4100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..18
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     103..122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        103..121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             205..227
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             247..264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             268..300
                     /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(304..312,313..327)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             349..405
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1101..1120
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1650..1668)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1664..1711
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1808..2263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1895..1914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2105..2126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2289..2393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2394..3188
                     /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             2394..3188
                     /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(2448..2467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3058..3077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3359..3947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3848..3867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
       61 cgttggccga ttcattaatg caggatctcg atcccgcgaa attaatacga ctcactatag
      121 ggagaccaca acggtttccc tctagaatca ccgagtttat tcttgacacc tgatgcgatg
      181 aatgatataa taggaaagta ctgttttgtt taactttaag aaggagatat acatatgcgg
      241 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
      301 cgggatctgt acgacgatga cgataaggat ccgatggtcg actcatcacg tcgtaagtgg
      361 aataaggcag gtcacgcagt cagagctata ggtcggctga gctcacgttg ggtttccgag
      421 tggatgtacc ccgaggacgg cgccctgaag agcgtgatca aggaggggtt gaggctgaag
      481 gacggcggcc actacgccgc cgaggtcagg accacctaca aggccaaaaa gcccgtgcag
      541 ctgcccggcg cctacatcgt cgacatcaag ttggacatcg tgtcccacaa cgaggactac
      601 accatcgtgg aacagtgcga acgcgccgag ggccgccacc ccaccggcgg catggtcggg
      661 ctgtacaagg gaggtacagg cgggagtctg gtgagcaagg gcgaggagga taacatggcc
      721 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      781 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg
      841 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtccct tcagttcatg
      901 tacggctcca aggcctacat taagcaccca gccgacatcc ccgactactt caagctgtcc
      961 ttccccgagg gcttcaggtg ggagcgcgtg atgatcttcg aggacggcgg cattattcac
     1021 gttaaccagg actcctccct gcaggacggc gtattcatct acaaggtgaa gctgcgcggc
     1081 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggagcctacg
     1141 cgtgaccaac tgactgaaga gcagatcgca gagtttaaag aggctttctc cctatttgac
     1201 aaggacgggg atgggacgat aacaaccaag gagctgggga cggtgttgcg gtctctgggg
     1261 cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac
     1321 ggcacattcg acttccctga gttcctgacg atgatggcaa ggaaaatgaa tgactcagac
     1381 agtgaagagg aaattagaga agcgttccgc gtgtttgata aggacggcaa tggctacatc
     1441 ggcgcagcag agcttcgcca cgtgatgaca gaccttggag agaagttaac agatgaggag
     1501 gttgatgaaa tgatcagggt agcagacatc gatggggatg gtcaggtaaa ctacgaagag
     1561 tttgtacaaa tgatgacagc gaagtaggaa ttcgaagctt gatccggctg ctaacaaagc
     1621 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1681 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg
     1741 cgtaatagcg aagaggccag caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1801 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1861 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1921 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1981 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2041 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2101 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2161 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2221 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     2281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgattg
     2401 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2461 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2521 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     2581 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2641 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2701 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2761 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2821 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2881 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     2941 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3001 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3061 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3121 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3181 tcttctgact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3241 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3301 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3361 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3421 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3481 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3541 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3601 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3661 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3721 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3781 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3841 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3901 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3961 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4021 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4081 gcagccgaac gaccgagcgc
//
