LOCUS       sequence-221536-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of orange fluorescent Ca2+ sensor O-GECO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5318)
  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 5318)
  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: pBAD-O-GECO1
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..54
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             58..90
                     /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(94..102,103..117)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             148..204
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWIKAGHAVRAIGRLSS"
     primer_bind     900..919
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1425..1442)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1595..1681
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1773..1800
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1819..1910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1911..1979,1980..2771)
                     /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(2129..2148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2942..3530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3431..3450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3684..3701
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3716..3856
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3782..3801)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3942..3964
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4120..4998)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5025..5309
                     /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     5232..5251
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 ttgggctaac aggaggaatt aaccatgggg ggttctcatc atcatcatca tcatggtatg
       61 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
      121 cctggctcga gcatggtcga ctcttcacgt cgtaagtgga ttaaggcagg tcacgcagtc
      181 agagctatag gtcggctgag ctcacccgtg gtttccgagc ggatgtaccc cgaggacggc
      241 gtcctgaaga gcgagatcaa gaaggggctg aggctgaagg acggcggcca ctacgccgcc
      301 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
      361 gacataaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga acagtgcgaa
      421 cgcgccgagg gccgccaccc caccggcggc agggacgagc tgtacaaggg aggtacagga
      481 gggagtctgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      541 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      601 gagggccgcc cctacgaggc ctttcagacc gctaagctga aggtgaccaa gggtggcccc
      661 ctgcccttcg cctgggacat cctgtcccct cagttcacat acggctccaa ggcctacatt
      721 aagcacccag ccgacatccc cgactacttt aagctgtcct tccccgaggg cttcaggtgg
      781 gagcgcgtga tgaacttcga ggacggcggc attattcacg ttaaccagga ctcctccctg
      841 caggacggcg tattcatcta caaggtgaag ctgcgcggca ccaacttccc ccccgacggc
      901 cccgtaatgc agaagaagac catgggctgg gaggctacgc gtgaccaact gactgaagag
      961 cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacgata
     1021 acaaccaagg agctggggac ggtgttgcgg tctctggggc agaaccccac agaagcagag
     1081 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacattcga cttccctgag
     1141 ttcctgacga tgatggcaag aagaatgaat gacacagaca gtgaagtgga aattagagaa
     1201 gcgttccgcg tgtttgataa tgacggcaat ggctacatcg gcgcagcaga gcttcgccac
     1261 gtgatgacag accttggaga gaagttaaca gatgaggagg ttgatgaaat gatcagggta
     1321 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     1381 aagtagaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     1441 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     1501 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     1561 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     1621 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     1681 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     1741 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     1801 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1861 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1921 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1981 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2041 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2101 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2161 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2221 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2281 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2341 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2401 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2461 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2521 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2581 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2641 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2701 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2761 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2821 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3061 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3541 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3601 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3661 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3721 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     3781 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3841 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3901 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3961 gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc
     4021 gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg
     4081 ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac
     4141 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     4201 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     4261 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     4321 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     4381 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     4441 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     4501 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     4561 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     4621 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     4681 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     4741 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     4801 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     4861 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     4921 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     4981 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     5041 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     5101 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     5161 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     5221 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     5281 tttatcgcaa ctctctactg tttctccata cccgtttt
//
