LOCUS       sequence-235117-        6620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mercury sensor with an optimized HrpRS-based amplifier. pSB3K3 
            carrying J109-32merR-t-PmerT-30hrpR-30hrpS-t-PhrpLE-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  AUTHORS   Wan X, Volpetti F, Petrova E, French C, Maerkl SJ, Wang B
  TITLE     Cascaded amplifying circuits enable ultrasensitive cellular sensors 
            for toxic metals.
  JOURNAL   Nat Chem Biol. 2019 May;15(5):540-548. doi: 
            10.1038/s41589-019-0244-3. Epub 2019 Mar 25.
  PUBMED    30911179
REFERENCE   2  (bases 1 to 6620)
  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: 
            "10.1038/s41589-019-0244-3"; journalName: "Nat Chem Biol"; date: 
            "2019-05"; volume: "15"; issue: "5"; pages: "540-548"
COMMENT     Sequence Label: pXWJ109HgAmp30E
FEATURES             Location/Qualifiers
     source          1..6620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        23..57
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     terminator      536..607
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      623..650
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             733..744
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1655..1666
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             1729..1740
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2670..2741
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2757..2784
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3018..3029
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3036..3749
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3066..3094)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3689..3709
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3772..3843
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3859..3886
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3893..3913
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(4131..4946)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4131..4946)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4854..4873
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4938..4957
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5541..6085)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5621..5640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6553..6571
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6575..6618)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agtttacagc tagctcagtc ctagggactg tgctagctac
       61 tagagtcaca caggaaagta ctagatggaa aataatttgg aaaacctgac cattggcgtt
      121 tttgccaagg cggccggggt caacgtggag acaatccgct tctatcagcg caagggcctg
      181 ttgcgggaac cggacaagcc ttacggcagc atccgccgct atggggaggc ggacgtggtt
      241 cgggtgaaat tcgtgaaatc ggcacagcgg ctggggttca gtctggacga gattgccgag
      301 ctgttgcggc tcgacgatgg cacccactgc gaggaggcca gcagcctggc cgaacacaag
      361 ctcaaggacg tgcgcgagaa gatggccgac ttggcgcgca tggaaaccgt gctgtctgaa
      421 ctcgtgtgcg cctgccatgc acgaaagggg aatgtttcct gcccgttgat cgcgtcacta
      481 cagggcgaag caggcctggc aaggtcagct atgccttagt actagagcca ggcatcaaat
      541 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
      601 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatgga
      661 tccgttccat atcgcttgac tacgtacatg agtacggaag taaggttacg ctatccaatc
      721 ctactagaga ttaaagagga gaaatactag atgagtacag gcatcgataa ggacgtccga
      781 gagtgttggg gcgtaactgc attatcagcg ggtcatcaaa ttgcaatgaa tagcgcgttt
      841 ctggatatgg acttgctgtt gtgcggggaa accggcaccg gcaaggacac actggccaac
      901 cgcattcacg agttgtccag caggtcggga ccctttgtgg gcatgaactg cgccgccatt
      961 cccgagtcgc tggcagagag ccagttattc ggtgtggtca acggtgcatt caccggcgta
     1021 tgccgggctc gcgagggcta catagaggcc tccagtggtg gcaccttgta cctggatgaa
     1081 atcgacagca tgccgttgag cctgcaagcc aaactgctgc gtgtgttgga gagtcgaggt
     1141 atcgagcgtc tgggctcgac cgaatttatc ccggtggatc tgcggatcat tgcctcggcc
     1201 cagcggccac tggatgaact ggtggaacaa ggacttttcc gtcgcgacct gttttttcgg
     1261 ctcaacgtgc tgacgcttca cttgccagcc ttgcgcaaac gtcgtgaaca gatcctgcca
     1321 ttgttcgacc agttcaccca gggtatcgct gccgagttcg gacgtcccgc tcctgcgctg
     1381 gacagcgggc gtgtgcagct gctgctcagc cacgactggc cgggcaacat ccgcgaattg
     1441 aagtctgcgg ccaagcgctt cgtactcggc ttccccttgc tgggcgccga ccctgtggaa
     1501 gcgcttgacc ctgccacggg gctgcgcacg caaatgcgca tcatcgagaa aatgctcatc
     1561 caggatgcct tgaagcggca caggcacaat ttcgacgcgg tgcttcagga gttggagttg
     1621 ccaagacgca ccctgtatca ccgcatgaag gaactgggag ttgcagcgcc gatcgctgcg
     1681 acggccgggg tctaataaaa cgcaggtgtg gttatgctac tagagattaa agaggagaaa
     1741 tactagatga tgagtcttga tgaaaggttt gaggatgatc tggacgagga gcgggttccg
     1801 aatctgggga tagttgccga aagtatttcg caactgggta tcgacgtgct gctatcgggt
     1861 gagaccggca cgggcaaaga cacgattgcc cgacggattc atgagatgtc aggccgcaaa
     1921 gggcgcctgg tggcgatgaa ttgcgcggcc attccggagt ccctcgccga gagcgagtta
     1981 ttcggcgtgg tcagcggtgc ctacaccggc gctgatcgct ccagagtcgg ttatgtcgaa
     2041 gcggcgcagg gcggcacgct gtacctggat gagatcgata gcatgccgct gagcctgcaa
     2101 gccaaattgc tgagggtgct ggaaacccga gcgcttgaac ggctgggttc gacgtcgacg
     2161 atcaagctgg atatctgcgt gatcgcctcc gcccaatgct cgctggacga cgccgtcgag
     2221 cgggggcagt ttcgtcgcga tctgtatttt cgcctgaacg tcctgacact caagcttcct
     2281 ccgctacgta accagtctga tcgcatagtt cccctgttca cacgttttac ggccgccgcc
     2341 gcgagggagc tcggtgttcc cgttcccgat gtttgcccac tgctgcacaa agtgctgctg
     2401 ggccacgact ggcccggcaa tatccgtgag ctcaaggcgg cagccaaacg ccatgtgctg
     2461 ggtttcccct tgctgggcgc cgagccgcag ggcgaagagc acttggcctg tgggctcaaa
     2521 tcgcaattgc gagtgatcga aaaagccctg attcaggagt cgctcaagcg ccacgacaat
     2581 tgtgtggatt cggtaagcct ggaactggac gtgccacgcc gtacgctcta tcgacgcatc
     2641 aaagaattgc agatctaata accaggcatc aaataaaacg aaaggctcag tcgaaagact
     2701 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     2761 caccttcggg tgggcctttc tgcgtttata tactagaggc cggattatgt ccgctgagtg
     2821 ggtcacggtc ccggatcagt tcccttgcga agctgaccga tgtttttgtg ccaaaagctg
     2881 ttgtggcaaa aaacggtttg cgcaaagttt tgtattacaa agaatttcac attttaaaat
     2941 atctttataa atcaatcagt tatttctatt ttcaagctgg cacggttatt gctatagggc
     3001 ttgtactact agagattaaa gaggagaaat actagatgcg taaaggagaa gaacttttca
     3061 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     3121 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
     3181 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc
     3241 aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc
     3301 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     3361 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     3421 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
     3481 atgtatacat catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac
     3541 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     3601 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     3661 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     3721 ttacacatgg catggatgaa ctatacaaat aataatacta gagccaggca tcaaataaaa
     3781 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     3841 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagta
     3901 gcggccgctg cagtccggca aaaaaacggg caaggtgtca ccaccctgcc ctttttcttt
     3961 aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact cgctgcgctc
     4021 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatct cgagtcccgt
     4081 caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac
     4141 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     4201 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     4261 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     4321 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     4381 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     4441 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     4501 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     4561 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     4621 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     4681 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     4741 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     4801 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     4861 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa
     4921 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     4981 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     5041 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     5101 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     5161 tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg
     5221 gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctag
     5281 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
     5341 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
     5401 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     5461 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     5521 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     5581 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5641 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     5701 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     5761 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     5821 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     5881 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     5941 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     6001 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     6061 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg acgctcagtg
     6121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6481 ctccatccag tctattccat ggtgccacct gacgtctaag aaaccattat tatcatgaca
     6541 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     6601 ttcgctaagg atgatttctg
//
