LOCUS       sequence-372329-        6618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded calcium indicator 
            SCW-HHL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6618)
  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 6618)
  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-HHL
FEATURES             Location/Qualifiers
     source          1..6618
                     /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)"
     CDS             1354..1371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1375..1407
                     /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             1420..1440
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2605..2661
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(3400..3417)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3570..3656
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3749..3776
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3795..3886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3887..3955,3956..4747)
                     /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(4105..4124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4789..5244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4876..4895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5086..5107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5355..5943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5844..5863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6097..6114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6129..6269
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6195..6214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6355..6377
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6515..6534
                     /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 tatgcggggt tctcatcatc atcatcatca tggtatggct
     1381 agcatgactg gtggacagca aatgggtcgg gatctgtacg agaacctgta cttccagggc
     1441 tcgagaatgg tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac
     1501 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1561 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc
     1621 tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcatcaa gcaccccgcc
     1681 gacatccccg actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg
     1741 aacttcgagg acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc
     1801 ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag
     1861 aagaagacaa tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg
     1921 aagggcgaca ttcatatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc
     1981 aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc
     2041 aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc
     2101 gagggccgcc accacctagg cggcgcatgc ctgccggacc aactgactga agagcagatc
     2161 gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc
     2221 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     2281 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     2341 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     2401 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     2461 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     2521 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagggt
     2581 ggcggaggtt ctgtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga
     2641 gctataggtc ggctgagctc actagagggc gaggcattga tcaaggagta catgcggttc
     2701 aaggtgcaca tggagggctc catggacggc cacgagttcg agatcgaggg cgagggcgag
     2761 ggccgcccct acgagggcac ccataccgcc aagctgaagg tgaccaaggg tggccccctg
     2821 cccttctcct gggacatcct gtcccctcag ttcgggtacg gctccagggc cttcatcaag
     2881 caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt caagtgggag
     2941 cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac ctccctggag
     3001 gacggcaccc tgatccacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc
     3061 gtaatgcaga ggaagacaat gggctgggaa gcgtccaccg agcggttgta ccccgaggac
     3121 ggcgtgctga agggcgacat tcatatggcc ctgcgcctga aggacggcgg ccgctacctg
     3181 gcggactgca agaccaccta caaggccaag aagcccgtgc agatgcccgg cgcctacaac
     3241 gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt ggaacagtac
     3301 gaacgctcag agggccgcca ccacctaggc ggcatggacg agctgtacaa gtaagaattc
     3361 gaagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     3421 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     3481 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     3541 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     3601 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     3661 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gntggcgggc aggacgcccg
     3721 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     3781 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3841 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3901 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3961 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4021 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4081 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     4141 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4201 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4261 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4321 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4381 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     4441 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4501 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4561 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4621 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4681 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4741 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
     4801 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4861 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4921 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4981 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5041 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5101 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     5161 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5221 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
     5281 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5341 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5401 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5461 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5521 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5581 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5641 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5701 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5761 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5821 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5881 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5941 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6001 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6061 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6121 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatc
     6181 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     6241 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     6301 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6361 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag aaggagatgg
     6421 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     6481 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     6541 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct
     6601 catgtttgac agcttatc
//
