LOCUS       sequence-372388-        6182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded protease indicator SPW.
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
FEATURES             Location/Qualifiers
     source          1..6182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     172..189
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    204..344
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(270..289)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     430..452
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     590..609
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(789..1667)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1694..1978
                     /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     1901..1920
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2005..2027
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2047..2064
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2068..2100
                     /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             2113..2133
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2866..2877
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3658..3675)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3828..3914
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4006..4033
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4052..4143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4144..4212,4213..5004)
                     /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(4362..4381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5046..5501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5133..5152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5343..5364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      join(5612..6182,1..18)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6101..6120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
       61 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      121 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      181 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      241 atttcacacc gcatctggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      301 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
      361 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
      421 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
      481 ggcagaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
      541 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
      601 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
      661 cgtagaggat ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg
      721 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
      781 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
      841 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
      901 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
      961 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     1021 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     1081 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     1141 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     1201 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     1261 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     1321 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     1381 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     1441 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     1501 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     1561 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     1621 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     1681 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     1741 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     1801 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     1861 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     1921 ataagattag cggatactac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     1981 ccgttttttt gggctagaaa taattttgtt taactttaag aaggagatat acatatgcgg
     2041 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
     2101 cgggatctgt acgagaacct gtacttccag ggctcgagaa tggtgagcaa gggcgaggca
     2161 gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa cagccacgag
     2221 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2281 aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg
     2341 tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc
     2401 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc
     2461 gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc
     2521 accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc
     2581 accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc
     2641 ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc
     2701 gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag
     2761 gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcggt
     2821 accggaggga gtgggggaac aggcgggagt ggaggaacag ggggatggga attgcaaagc
     2881 gggggcactg gcgggtcggg tggtactggt ggaagtggtg catgcggcga ggcattgatc
     2941 aaggagtaca tgcggttcaa ggtgcacatg gagggctcca tggacggcca cgagttcgag
     3001 atcgagggcg agggcgaggg ccgcccctac gagggcaccc ataccgccaa gctgaaggtg
     3061 accaagggtg gccccctgcc cttctcctgg gacatcctgt cccctcagtt cgggtacggc
     3121 tccagggcct tcatcaagca ccccgccgac atccccgact actataagca gtccttcccc
     3181 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc
     3241 caggacacct ccctggagga cggcaccctg atccacaagg tgaagctccg cggcaccaac
     3301 ttccctcctg acggccccgt aatgcagagg aagacaatgg gctgggaagc gtccaccgag
     3361 cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta agatggccct gcgcctgaag
     3421 gacggcggcc gctacctggc ggactgcaag accacctaca aggccaagaa gcccgtgcag
     3481 atgcccggcg cctacaacgt cgaccgcaag ttggacatca cctcccacaa cgaggactac
     3541 accgtggtgg aacagtacga acgctcagag ggccgccact ccaccggcgg catggacgag
     3601 ctgtacaagt aagaattcga agcttggctg ttttggcgga tgagagaaga ttttcagcct
     3661 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     3721 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     3781 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     3841 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     3901 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     3961 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     4021 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     4081 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4141 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4201 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4261 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4321 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4381 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4441 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4501 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4561 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4621 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4681 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4741 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4801 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4861 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4921 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4981 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5041 gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5101 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5161 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5221 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5281 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5341 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     5401 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5461 atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga
     5521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6181 gg
//
