LOCUS       sequence-372842-        6182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded protease indicator 
            SPW-HHL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6182)
  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 6182)
  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-SPW-HHL
FEATURES             Location/Qualifiers
     source          1..6182
                     /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             2173..2184
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2965..2982)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3135..3221
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3313..3340
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3359..3450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3451..3519,3520..4311)
                     /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(3669..3688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4353..4808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4440..4459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4650..4671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4919..5507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5408..5427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5661..5678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5693..5833
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5759..5778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5919..5941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6079..6098
                     /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 cggcggtacc ggagggagtg ggggaacagg cgggagtgga
     2161 ggaacagggg gatgggaatt gcaaagcggg ggcactggcg ggtcgggtgg tactggtgga
     2221 agtggtgcat gcggcgaggc attgatcaag gagtacatgc ggttcaaggt gcacatggag
     2281 ggctccatgg acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2341 ggcacccata ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
     2401 atcctgtccc ctcagttcgg gtacggctcc agggccttca tcaagcaccc cgccgacatc
     2461 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2521 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
     2581 cacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaggaag
     2641 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
     2701 gacattcata tggccctgcg cctgaaggac ggcggccgct acctggcgga ctgcaagacc
     2761 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     2821 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctcagagggc
     2881 cgccaccacc taggcggcat ggacgagctg tacaagtaag aattcgaagc ttggctgttt
     2941 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     3001 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     3061 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     3121 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     3181 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     3241 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     3301 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3481 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3541 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3601 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3661 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     3721 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3781 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3841 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3901 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3961 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     4021 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4081 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4141 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4201 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4261 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4321 caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc
     4381 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4441 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4501 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4561 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4621 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4681 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4741 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4801 tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4861 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4921 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4981 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5041 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5101 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5161 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5221 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5281 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5341 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5401 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5461 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5521 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5581 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5641 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5701 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catctggtgc actctcagta
     5761 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     5821 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5881 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5941 gttttcaccg tcatcaccga aacgcgcgag gcagaaggag atggcgccca acagtccccc
     6001 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6061 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6121 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     6181 tc
//
