LOCUS       sequence-372336-        6179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded protease indicator 
            SPW-RHH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  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 6179)
  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-RHH
FEATURES             Location/Qualifiers
     source          1..6179
                     /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(2962..2979)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3132..3218
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3310..3337
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3356..3447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3448..3516,3517..4308)
                     /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(3666..3685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4350..4805
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4437..4456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4647..4668
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4916..5504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5405..5424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5658..5675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5690..5830
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5756..5775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5916..5938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6076..6095
                     /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 accccaggga cggcgtgctg
     1921 aagggcgaca ttcatatggc cctgcgcctg aaggacggcg gccactacct ggcggacttc
     1981 aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc
     2041 aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc
     2101 gagggccgcc actccaccgg 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 ttgtacccca gggacggcgt gctgaagggc
     2701 gacattcata tggccctgcg cctgaaggac ggcggccact acctggcgga ctgcaagacc
     2761 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     2821 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctcagagggc
     2881 cgccactcca ccggcggcat ggacgagctg tactaagaat tcgaagcttg gctgttttgg
     2941 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     3001 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     3061 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     3121 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     3181 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3241 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3301 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     3361 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3421 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3481 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3541 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3601 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3661 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     3721 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3781 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3841 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3901 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3961 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4021 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4081 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4141 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4201 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4261 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4321 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     4381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4501 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4561 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4621 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4681 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4741 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4801 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5041 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5581 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5641 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     5701 attttctcct tacgcatctg tgcggtattt cacaccgcat ctggtgcact ctcagtacaa
     5761 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     5821 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5881 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5941 ttcaccgtca tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc
     6001 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6061 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6121 ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatc
//
