LOCUS       sequence-338854-        5813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bioluminescent Red Indicator for Calcium (bacterial expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5813)
  AUTHORS   Tian X, Zhang Y, Li X, Xiong Y, Wu T, Ai HW
  TITLE     A luciferase prosubstrate and a red bioluminescent calcium indicator
            for imaging neuronal activity in mice.
  JOURNAL   Nat Commun. 2022 Jul 8;13(1):3967. doi: 10.1038/s41467-022-31673-x.
  PUBMED    35803917
REFERENCE   2  (bases 1 to 5813)
  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"; journalName: "Nat 
            Commun."; date: "2022-07-8"; volume: "13(1):3967. doi"; pages: " 
            10.1038/s41467-022-31673-x"
COMMENT     Sequence Label: pBAD-BRIC
FEATURES             Location/Qualifiers
     source          1..5813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(560..578)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     828..847
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1951..2028
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2149..2172
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2214..2231)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2384..2470
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2562..2589
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2608..2699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2700..2768,2769..3560)
                     /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(2918..2937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3731..4319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4220..4239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4473..4490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4505..4645
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4571..4590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4731..4753
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4909..5787)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagcatgg tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac
      481 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      541 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc
      601 tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcatcaa gcaccccgcc
      661 gacatccccg actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg
      721 aacttcgagg acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc
      781 ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag
      841 aagaagacaa tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg
      901 aagggcgaca ttaagatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc
      961 aagaccatct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc
     1021 aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc
     1081 gagggccgcc atcttgatac actcgaagat ttcgttgggg actggcgaca gacagccggc
     1141 tacaacttga gtcaaatcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg
     1201 gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac
     1261 atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa
     1321 atttttaagg tggtgtaccc tgtggataat catcatttta aggtgatcct gcactatggc
     1381 acactggtaa tcgacggggt tacgccgaac ttgatcgact atttcggacg gccgtatgaa
     1441 ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgat tatgcatgac
     1501 caactgacag aagagcagat tgcagagttc aaagaagcct tctcattatt cgacaaggat
     1561 ggggacggca ccatcaccac aaaggaactt ggtaccgtta tgaggtcgct tggacaaaac
     1621 ccaacggaag cagaattgca ggatatgatc aatgaagtcg atgctgatgg caatggaacg
     1681 atttactttc ctgaatttct tactatgatg gctagaaaaa tgaaggacac agacagcgaa
     1741 gaggaaatcc gagaagcatt ccgtgttttt gacaaggatg ggaacggcta catcagcgct
     1801 gctcaactac gtcacgtcat gacaaacctt ggggagaagt taacagatga agaagttgat
     1861 gaaatgataa gggaagcaga tatcgatggt gatggccaag taaactatga agagtttgta
     1921 caaatgatga cagcaaaggg gggcgagtcc aagaggcgct ggaagaaaaa cttcattgcc
     1981 gtcagcgctg ccaaccggtt caagaagatc tccagctccg gggcactgga gctctggaaa
     2041 ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta
     2101 accatcaacg gagtgaccgg ctggcgtctg catgaacgca ttctggcgtg gagccacccg
     2161 cagttcgaaa aataaaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     2221 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     2281 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     2341 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     2401 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     2461 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     2521 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     2581 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     2641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2881 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2941 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3601 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3841 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4321 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4381 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4441 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4501 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4621 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4681 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4741 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     4801 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     4861 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     4921 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     4981 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     5041 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     5101 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     5161 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     5221 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     5281 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     5341 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     5401 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     5461 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     5521 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     5581 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     5641 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     5701 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     5761 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
