LOCUS       sequence-373780-        6542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded potassium indicator SKW.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6542)
  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 6542)
  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-SKW
FEATURES             Location/Qualifiers
     source          1..6542
                     /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"
     primer_bind     complement(3325..3342)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3495..3581
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3673..3700
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3719..3810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3811..3879,3880..4671)
                     /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(4029..4048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4713..5168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4800..4819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5010..5031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5279..5867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5768..5787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6021..6038
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6053..6193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6119..6138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6279..6301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6439..6458
                     /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 ttaagatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc
     1981 aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc
     2041 aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc
     2101 gagggccgcc actccaccgg cggcgcatgc ctgccggacg gcctgttcaa cttcgtgaag
     2161 gacgccggcg agaagctgtg ggatgccgtg acaggccagc acgacaagga cgaccaggcc
     2221 aagaaggtgc aggaacacct gaacaagacc ggcatccccg acgccgacaa agtgaacatc
     2281 cagatcgccg acggcaaggc cacagtgaca ggcgacggac tgagccagga agccaaagaa
     2341 aagatcctgg tggccgtggg caacatcagc ggaatcgcca gcgtggacga tcaagtgaaa
     2401 accgccaccc ctgccaccgc cagccagttc tacacagtga agtccggcga caccctgagc
     2461 gccatcagca aacaggtgta cggcaacgcc aacctgtaca acaagatttt cgaggccaac
     2521 aagcccatgc tgaagtcccc cgacaaaatc taccccggcc aggtgctgag aatccccgag
     2581 agctcactag agggcgaggc attgatcaag gagtacatgc ggttcaaggt gcacatggag
     2641 ggctccatgg acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2701 ggcacccata ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
     2761 atcctgtccc ctcagttcgg gtacggctcc agggccttca tcaagcaccc cgccgacatc
     2821 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2881 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
     2941 cacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaggaag
     3001 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
     3061 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga ctgcaagacc
     3121 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     3181 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctcagagggc
     3241 cgccactcca ccggcggcat ggacgagctg tacaagtaag aattcgaagc ttggctgttt
     3301 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     3361 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     3421 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     3481 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     3541 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     3601 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     3661 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     3721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4021 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     4081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4321 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     4381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4681 caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc
     4741 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4801 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4861 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4921 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4981 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5041 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5101 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     5161 tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5221 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5281 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5341 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5401 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5461 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5521 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5581 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5641 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5701 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5761 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5821 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5881 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5941 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6001 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     6061 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catctggtgc actctcagta
     6121 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     6181 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6241 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6301 gttttcaccg tcatcaccga aacgcgcgag gcagaaggag atggcgccca acagtccccc
     6361 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6421 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6481 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     6541 tc
//
