LOCUS       sequence-221540-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of red fluorescent Ca2+ sensor R-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-R-GECO1
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             47..64
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             68..100
                     /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(104..112,113..127)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             158..214
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     910..929
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             953..1396
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(1435..1452)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1605..1691
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1783..1810
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1829..1920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1921..1989,1990..2781)
                     /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(2139..2158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2952..3540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3441..3460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3694..3711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3726..3866
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3792..3811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3952..3974
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4130..5008)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5035..5318,1)
                     /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     5242..5261
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 tacccgtttt ttgggctaac aggaggaatt aaccatgggg ggttctcatc atcatcatca
       61 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
      121 cgataaggat cctggctcga gcatggtcga ctcatcacgt cgtaagtgga ataaggcagg
      181 tcacgcagtc agagctatag gtcggctgag ctcacccgtg gtttccgagc ggatgtaccc
      241 cgaggacggc gccctgaaga gcgagatcaa gaaggggctg aggctgaagg acggcggcca
      301 ctacgccgcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
      361 ctacatcgtc gacatcaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga
      421 acagtgcgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg
      481 aggtacaggc gggagtctgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
      541 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      601 gggcgagggc gagggccgcc cctacgaggc ctttcagacc gctaagctga aggtgaccaa
      661 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
      721 ggcctacatt aagcacccag ccgacatccc cgactacttc aagctgtcct tccccgaggg
      781 cttcaggtgg gagcgcgtga tgaacttcga ggacggcggc attattcacg ttaaccagga
      841 ctcctccctg caggacggcg tattcatcta caaggtgaag ctgcgcggca ccaacttccc
      901 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggctacgc gtgaccaact
      961 gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga
     1021 tgggacgata acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac
     1081 agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacattcga
     1141 cttccctgag ttcctgacga tgatggcaag aaaaatgaat gacacagaca gtgaagagga
     1201 aattagagaa gcgttccgcg tgtttgataa ggacggcaat ggctacatcg gcgcagcaga
     1261 gcttcgccac gtgatgacag accttggaga gaagttaaca gatgaggagg ttgatgaaat
     1321 gatcagggta gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat
     1381 gatgacagcg aagtagaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     1441 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     1501 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1561 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1621 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1681 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1741 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1801 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     1861 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1921 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1981 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2041 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2101 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2161 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2221 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2281 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2341 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2401 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2461 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2521 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2581 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2641 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2701 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2761 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2821 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2881 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2941 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3001 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3061 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3121 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3181 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3241 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3301 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3361 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3421 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3481 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3541 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3601 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3661 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3721 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3781 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3841 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3901 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3961 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
     4021 gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     4081 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     4141 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     4201 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     4261 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     4321 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     4381 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     4441 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     4501 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     4561 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     4621 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     4681 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     4741 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     4801 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     4861 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     4921 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     4981 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     5041 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     5101 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     5161 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     5221 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta
     5281 cctgacgctt tttatcgcaa ctctctactg tttctcca
//
