LOCUS       sequence-221539-        5477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of red fluorescent Ca2+ sensor jREX-GECO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5477)
  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 5477)
  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: pTorPE-jREX-GECO1
FEATURES             Location/Qualifiers
     source          1..5477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     497..516
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1022..1039)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1192..1278
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1370..1397
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1416..1507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1508..1576,1577..2368)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1726..1745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2539..3127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3028..3047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3281..3298
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3313..3453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3379..3398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3539..3561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3717..4595)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4622..4906
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4829..4848
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4948..4959
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5114..5131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5135..5167
                     /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(5171..5179,5180..5194)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             5222..5278
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
ORIGIN
        1 tcgtggaaca gtgcgaacgc gccgagggcc gccaccccac cggcggcatg gtcgggctgt
       61 acaagggagg tacaggcggg agtctggtga gcaagggcga ggaggataac atggccatca
      121 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      181 agatcgaggg cgagggcgag ggccgcccct acgaggcctt tcagaccgct aagctgaagg
      241 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccttcag ttcatgtacg
      301 gctccaaggc ctacattaag cacccagccg acatccccga ctacttcaag ctgtccttcc
      361 ccgagggctt caggtgggag cgcgtgatga tcttcgagga cggcggcatt attcacgtta
      421 accaggactc ctccctgcag gacggcgtat tcatctacaa ggtgaagctg cgcggcacca
      481 acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag cctacgcgtg
      541 acgatctgac tgaagagcag atcgcagagt ttaaagaggc tttctcccta tttgacaagg
      601 acggggatgg gacgataaca accaaggagc tggggacggt gttccggtct ctggggcaga
      661 accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac ggtgacggca
      721 cattcgactt ccctgagttc ctgacgatga tggcaaggaa aatgaatgac tcagacagtg
      781 aagaggaaat tagagaagcg ttccgcgtgt ttgataagga cggcaatggc tacatcggcg
      841 cagcagagct tcgccacgtg atgacagacc ttggagagaa gttaacagat gaggaggttg
      901 atgaaatgat cagggtagca gacatcgatg gggatggtca ggtaaactac gaagagtttg
      961 tacaaatgat gacagcgaag tagaagcttg gctgttttgg cggatgagag aagattttca
     1021 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1081 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1141 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1201 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1261 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     1321 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     1381 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     1441 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1501 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1561 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1621 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1681 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1741 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1801 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1861 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1921 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1981 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2041 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2101 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2161 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2221 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2281 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2341 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2401 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2461 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2521 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2581 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2641 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2701 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2761 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2821 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2881 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2941 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3001 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3061 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3121 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3181 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3241 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3301 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3361 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3421 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3481 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3541 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc
     3601 aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga cgaagcaggg
     3661 attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat
     3721 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
     3781 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
     3841 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
     3901 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
     3961 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
     4021 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
     4081 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     4141 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     4201 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     4261 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     4321 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     4381 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     4441 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     4501 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     4561 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     4621 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     4681 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     4741 aaagccatga caaaatcgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     4801 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     4861 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg
     4921 ctaacaggag gaattaacca tgggtttaaa gaggagaaag gtcatgaaca ataacgatct
     4981 ctttcaggca tcacgtcggc gttttctggc acaactcggc agcttaaccg acgccgggac
     5041 gctggggccg tcattgttaa cgccgcgacg tgcgactgcg gcgcaagcgg cgactgacgc
     5101 ttctagaggt tctcatcatc atcatcatca tggtatggct agcatgactg gaggacagca
     5161 aatgggtcgg gatctgtacg acgatgatga taaggatctc gccacaatgg ttgactcttc
     5221 acgtcgtaag tggaataagg caggtcacgc agtcagagct ataggtcggc tgagctcacg
     5281 ttgggtttcc gagtggatgt accccgagga cggcgccctg aagagcgtga tcaaggaggg
     5341 gttgaggctg aaggacggcg gccactacgc cgccgaggtc aggaccacct acaaggccaa
     5401 aaagcccgtg cagctgcccg gcgcctacat cgtcgacatc aagttggaca tcgtgtccca
     5461 caacgaggac tacacca
//
