LOCUS       sequence-240605-        4257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mercury sensor with optimized intracellular receptor density. 
            pSB3K3 carrying J109-32merR-t-PmerT-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4257)
  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 4257)
  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: pXWJ109Hg-gfp
FEATURES             Location/Qualifiers
     source          1..4257
                     /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             751..1464
                     /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(781..809)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1404..1424
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1487..1558
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1574..1601
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1608..1628
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(1768..2583)
                     /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(1768..2583)
                     /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     2491..2510
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2575..2594
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(3178..3722)
                     /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(3257..3276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4190..4208
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4212..4255)
                     /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 atgcgtaaag gagaagaact tttcactgga
      781 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
      841 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
      901 ggaaaactac ctgttccatg gccaacactt gtcactactt tcggttatgg tgttcaatgc
      961 tttgcgagat acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa
     1021 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
     1081 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     1141 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     1201 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
     1261 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
     1321 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     1381 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
     1441 catggcatgg atgaactata caaataataa tactagagcc aggcatcaaa taaaacgaaa
     1501 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     1561 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagtagcggc
     1621 cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1681 tatcagctca ctcaaaggcg gtaatctcga gtcccgtcaa gtcagcgtaa tgctctgcca
     1741 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     1801 caatttattc atatcaggat tatcgatacc atatttttga aaaagccgtt tctgtaatga
     1861 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1921 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1981 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     2041 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     2101 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     2161 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     2221 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     2281 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     2341 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     2401 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     2461 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     2521 agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct
     2581 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     2641 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     2701 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     2761 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     2821 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     2881 agcaacacct tcttcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat
     2941 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3001 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3061 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3121 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3181 ccataggctc cgccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
     3241 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
     3301 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
     3361 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
     3421 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3481 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
     3541 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
     3601 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc aaaaaaccgc
     3661 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
     3721 aagaagatca tcttattaag gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attccatggt
     4141 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4201 cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctg
//
