LOCUS       sequence-227506-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of red fluorescent Ca2+ sensor R-GECO1.2.
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-GECO1.2
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..298
                     /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     221..240
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             344..361
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             365..397
                     /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(401..409,410..424)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             455..511
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1207..1226
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1732..1749)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1902..1988
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2080..2107
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2126..2217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2218..2286,2287..3078)
                     /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(2436..2455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3249..3837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3738..3757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3991..4008
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4023..4163
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4089..4108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4249..4271
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4427..5305)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
       61 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
      121 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
      181 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
      241 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
      301 ccgttttttg ggctaacagg aggaattaac catggggggt tctcatcatc atcatcatca
      361 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      421 taaggatcct ggctcgagca tggtcgactc ttcacgtcgt aagtggaata aggcaggtca
      481 cgcagtcaga gctataggtc ggctgagctc acccgtggtt tccgagcgga tgtaccccga
      541 ggacggcgcc ctgaagagcg agatcaagaa ggggctgagg ctgaaggacg gcggccacta
      601 cgccgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
      661 catcgtcgac atcaagttgg acatcgtgtc ccacaacgag gactacacca tcgtggaaca
      721 gtgcgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagggagg
      781 tacaggcggg agtctggtga gcaagggcga ggaggataac agggccatcg tcaaggagtt
      841 catgcgcttc aagttgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      901 cgagggcgag ggccgcccct acgaggcctt tcagaccgct aagctgaagg tgaccaaggg
      961 tggccccctg cccttcgcat gggacatcct gtcccctcag ttgatgtacg gctccaaggc
     1021 ctacattaag cacccagccg acatccccga ctacttcaag ctgtccttcc ccgagggctt
     1081 caggtgggag cgcgtgatga acttcgagga cggcggcatt attcacgtta gccaggacac
     1141 ctccctgcag gacggcgtat tcatctacaa ggtgaagctg cgcggcacca acttcccccc
     1201 cgacggcccc gtaatgcaga agaagaccat gggctgggag gctacgcgtg accaactgac
     1261 tgaagagcag atcgcagaat ttaaagaggc tttctcccta tttgacaagg acggggatgg
     1321 gacgatgaca accaaggagc tggggacggt gatgcggtct ctggggcaga accccacaga
     1381 agcagagctg caggacatga tcaatgaagt agatgccgac ggtgacggca cattcgactt
     1441 ccctgagttc ctgacgatga tggcaagaaa aatgaatgac acagacagtg aagaggaaat
     1501 tagagaagcg ttccgcgtgt ttgataagga cggcaatggc tacatcggcg cagcagagct
     1561 tcgccacgtg atgacagacc ttggagagaa gataacagat gaggaggttg atgaaatgat
     1621 cagggtagca gacatcgatg gggatggtca ggtaaactat gaagagtttg tacaaatgat
     1681 gacagcgaag tagaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
     1741 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     1801 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     1861 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     1921 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1981 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     2041 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
     2101 gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat
     2161 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2221 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2281 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2341 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2401 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2461 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2521 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2581 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2641 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2701 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2761 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2821 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2881 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2941 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3001 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3061 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3361 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3901 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3961 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4021 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4081 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     4141 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     4201 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4261 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc aattcgcgcg
     4321 cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa
     4381 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga
     4441 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg
     4501 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga
     4561 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt
     4621 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg
     4681 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt
     4741 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact
     4801 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct
     4861 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat
     4921 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt
     4981 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc
     5041 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac
     5101 ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga
     5161 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt
     5221 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca
     5281 ggggatcatt ttgcgcttca gccatacttt tcatactc
//
