LOCUS       sequence-221532-        5291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of red fluorescent Ca2+ sensor jRCaMP1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5291)
  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 5291)
  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-jRCaMP1a
FEATURES             Location/Qualifiers
     source          1..5291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             53..70
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             74..106
                     /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(110..118,119..133)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             164..220
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(1414..1431)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1584..1670
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1762..1789
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1808..1899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1900..1968,1969..2760)
                     /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(2118..2137)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2931..3519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3420..3439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3673..3690
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3705..3845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3771..3790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3931..3953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4109..4987)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5014..5291,1..7)
                     /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     5221..5240
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 tctccatacc cgttttttgg gctaacagga ggaattaacc atggggggtt ctcatcatca
       61 tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga
      121 cgatgacgat aaggatccga gctcgagaat ggtcgactca tcacgtcgta agtggaataa
      181 gacaggtcac gcagtcagag ctataggtcg gctgagctca gcgatcaact gcgaaatgat
      241 gtacccagcg gatggtggtc tgcgtggtta cactcacatg gcgctgaaag ttgatggcgg
      301 cggtcacctg tcctgttctt tcgtgaccac ctaccgctcc aaaaagactg tcggcaacat
      361 taagatgcct gccattcata gcgtcagcca ccgtctggag cgcctggagg agagcgataa
      421 cgaaatgttt gtcgtacagc gtgaacacgc agttgccaag tttgtgggcc tgggtggtgg
      481 cggcggtacc ggagggagca tgaactccct gatcaaggag aacatgcgta tgaaagtggt
      541 tctggaaggc tccgtaaacg gccaccagtt caaatgcact ggtgaaggcg aaggcaaccc
      601 gtatatgggc acccagacta tgcgtatcaa agtgatcgag ggtggtccgc tgccgtttgc
      661 gttcgacatc ctggcgacgt cctttatgta tggctcccgt accttcatca aatatccgaa
      721 aggcatcccg gatttcttta agcagtcctt cccggaaggt tttacctggg aacgtgtgac
      781 ccgttacgaa gacggcggcg taattaccgt tatgcaagac acgtctctgg aggatggctg
      841 cctggtgtat cacgtgcagg ttcgcggtgt gaacttcccg agcaatggtg ctgtaatgca
      901 aaagaaaacc aaaggttggg agcctacgga ctcccaaacc actgaagagc agatcgcaga
      961 atttaaagag gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga
     1021 gctggggacg gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat
     1081 gatcaatgaa gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgattat
     1141 gatggcaaga aaaatgaaat acacagacag tgaagaagaa attagagaag cgttcggcgt
     1201 gtttgataag gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa
     1261 ccttggagag aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacagcga
     1321 tggggatggt caggtaaact acgaagagtt tgtacaaatg atgacagcga agtagaagct
     1381 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     1441 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     1501 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     1561 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1621 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     1681 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     1741 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     1801 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1861 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1921 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1981 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2041 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2101 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2161 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2221 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2281 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2341 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2401 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2461 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2521 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2581 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2641 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2701 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2761 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2821 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2881 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2941 ttttttctgc gcgtaatctg ctgcttgcaa acaaaacaac caccgctacc agcggtggtt
     3001 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3061 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3121 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3181 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3241 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3301 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3361 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3421 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3481 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3541 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3601 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3661 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3721 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3781 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3841 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
     4021 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
     4081 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
     4141 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     4201 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     4261 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     4321 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     4381 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     4441 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     4501 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     4561 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     4621 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     4681 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     4741 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     4801 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     4861 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     4921 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     4981 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
     5041 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     5101 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     5161 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     5221 atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac
     5281 tctctactgt t
//
