LOCUS       sequence-372786-        6673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded calcium indicator SCW.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6673)
  AUTHORS   Gohil K, Wu SY, Takahashi-Yamashiro K, Shen Y, Campbell RE
  TITLE     Biosensor Optimization Using a Forster Resonance Energy Transfer 
            Pair Based on mScarlet Red Fluorescent Protein and an 
            mScarlet-Derived Green Fluorescent Protein.
  JOURNAL   ACS Sens. 2023 Feb 24;8(2):587-597. doi: 10.1021/acssensors.2c01730.
            Epub 2023 Jan 24.
  PUBMED    36693235
REFERENCE   2  (bases 1 to 6673)
  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: 
            "10.1021/acssensors.2c01730"; journalName: "ACS Sens"; date: 
            "2023-02-24- 24"; volume: "8"; issue: "2"; pages: "587-597"
COMMENT     Sequence Label: pBAD-SCW
FEATURES             Location/Qualifiers
     source          1..6673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1312..1334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1368..1390
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1410..1427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1431..1463
                     /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             1476..1496
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2661..2717
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(3456..3473)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3626..3712
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3804..3831
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3850..3941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3942..4010,4011..4802)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4160..4179)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4844..5299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4931..4950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5141..5162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5410..5998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5899..5918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6152..6169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6184..6324
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6250..6269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6410..6432
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6570..6589
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atactacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagaaataa ttttgtttaa
     1321 ctttaagaag gagatataca tayscgkkkt tytsrkcwws aaataatttt gtttaacttt
     1381 aagaaggaga tatacatatg cggggttctc atcatcatca tcatcatggt atggctagca
     1441 tgactggtgg acagcaaatg ggtcgggatc tgtacgagaa cctgtacttc cagggctcga
     1501 gaatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     1561 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1621 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg
     1681 acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac cccgccgaca
     1741 tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1801 tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga
     1861 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     1921 agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg
     1981 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     2041 ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt
     2101 tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg
     2161 gccgccactc caccggcggc gcatgcctgc cggaccaact gactgaagag cagatcgcag
     2221 aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacaata acaaccaagg
     2281 agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca
     2341 tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa
     2401 tgatggcaag aaaaatgaaa tacagggaca cggaagaaga aattagagaa gcgttcggtg
     2461 tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac gtgatgacaa
     2521 accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa gcagacatcg
     2581 atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aagggtggcg
     2641 gaggttctgt cgactcatca cgtcgtaagt ggaataagac aggtcacgca gtcagagcta
     2701 taggtcggct gagctcacta gagggcgagg cattgatcaa ggagtacatg cggttcaagg
     2761 tgcacatgga gggctccatg gacggccacg agttcgagat cgagggcgag ggcgagggcc
     2821 gcccctacga gggcacccat accgccaagc tgaaggtgac caagggtggc cccctgccct
     2881 tctcctggga catcctgtcc cctcagttcg ggtacggctc cagggccttc atcaagcacc
     2941 ccgccgacat ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg
     3001 tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg
     3061 gcaccctgat ccacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa
     3121 tgcagaggaa gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg
     3181 tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg
     3241 actgcaagac cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg
     3301 accgcaagtt ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac
     3361 gctcagaggg ccgccactcc accggcggca tggacgagct gtacaagtaa gaattcgaag
     3421 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     3481 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     3541 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     3601 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3661 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     3721 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     3781 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     3841 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3901 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3961 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4021 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4081 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4141 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     4201 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4261 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4321 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4381 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4441 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     4501 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4561 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4621 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4681 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4741 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4801 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     4861 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4921 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4981 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5041 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5101 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5161 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5221 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5281 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5341 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5521 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5641 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5701 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5761 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5821 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5881 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5941 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6001 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6061 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6181 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatctggtg
     6241 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     6301 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     6361 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     6421 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc
     6481 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6541 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6601 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt
     6661 ttgacagctt atc
//
