LOCUS       sequence-243991-        7978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mercury sensor with two-layered amplifier cascade. pSB4A3 carrying 
            J109-32merR-t-PmerT-30hrpR-30hrpS-t-PhrpLE-30RinA-t-PrinA-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7978)
  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 7978)
  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: pXW109Hg(RS-RinA)
FEATURES             Location/Qualifiers
     source          1..7978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..59
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(83..100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(362..1312)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1360..1582
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2074..2168)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2198..2989,2990..3058))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2821..2840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3059..3163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3228..3246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3250..3293)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3296..3317
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        3318..3352
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     terminator      3831..3902
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3918..3945
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             4028..4039
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4950..4961
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             5024..5035
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      5970..6041
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6057..6084
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             6318..6329
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6775..6846
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6862..6889
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             7084..7095
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7102..7815
                     /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(7132..7160)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7755..7775
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      7838..7909
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7925..7952
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    join(7959..7978,1)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
       61 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatctcgag gttacattgt cgatctgttc
      181 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
      241 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
      301 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
      361 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
      421 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
      481 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
      541 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
      601 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
      661 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
      721 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
      781 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
      841 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
      901 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
      961 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     1021 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     1081 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     1141 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     1201 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     1261 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     1321 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     1381 atgtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac
     1441 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     1501 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     1561 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     1621 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     1681 ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg
     1741 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     1801 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     1861 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     1921 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     1981 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     2041 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     2101 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     2161 caggagtcca agcgagctcg taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt cccatggtgc
     3181 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3241 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt
     3301 cgcggccgct tctagagttt acagctagct cagtcctagg gactgtgcta gctactagag
     3361 tcacacagga aagtactaga tggaaaataa tttggaaaac ctgaccattg gcgtttttgc
     3421 caaggcggcc ggggtcaacg tggagacaat ccgcttctat cagcgcaagg gcctgttgcg
     3481 ggaaccggac aagccttacg gcagcatccg ccgctatggg gaggcggacg tggttcgggt
     3541 gaaattcgtg aaatcggcac agcggctggg gttcagtctg gacgagattg ccgagctgtt
     3601 gcggctcgac gatggcaccc actgcgagga ggccagcagc ctggccgaac acaagctcaa
     3661 ggacgtgcgc gagaagatgg ccgacttggc gcgcatggaa accgtgctgt ctgaactcgt
     3721 gtgcgcctgc catgcacgaa aggggaatgt ttcctgcccg ttgatcgcgt cactacaggg
     3781 cgaagcaggc ctggcaaggt cagctatgcc ttagtactag agccaggcat caaataaaac
     3841 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     3901 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atggatccgt
     3961 tccatatcgc ttgactacgt acatgagtac ggaagtaagg ttacgctatc caatcctact
     4021 agagattaaa gaggagaaat actagatgag tacaggcatc gataaggacg tccgagagtg
     4081 ttggggcgta actgcattat cagcgggtca tcaaattgca atgaatagcg cgtttctgga
     4141 tatggacttg ctgttgtgcg gggaaaccgg caccggcaag gacacactgg ccaaccgcat
     4201 tcacgagttg tccagcaggt cgggaccctt tgtgggcatg aactgcgccg ccattcccga
     4261 gtcgctggca gagagccagt tattcggtgt ggtcaacggt gcattcaccg gcgtatgccg
     4321 ggctcgcgag ggctacatag aggcctccag tggtggcacc ttgtacctgg atgaaatcga
     4381 cagcatgccg ttgagcctgc aagccaaact gctgcgtgtg ttggagagtc gaggtatcga
     4441 gcgtctgggc tcgaccgaat ttatcccggt ggatctgcgg atcattgcct cggcccagcg
     4501 gccactggat gaactggtgg aacaaggact tttccgtcgc gacctgtttt ttcggctcaa
     4561 cgtgctgacg cttcacttgc cagccttgcg caaacgtcgt gaacagatcc tgccattgtt
     4621 cgaccagttc acccagggta tcgctgccga gttcggacgt cccgctcctg cgctggacag
     4681 cgggcgtgtg cagctgctgc tcagccacga ctggccgggc aacatccgcg aattgaagtc
     4741 tgcggccaag cgcttcgtac tcggcttccc cttgctgggc gccgaccctg tggaagcgct
     4801 tgaccctgcc acggggctgc gcacgcaaat gcgcatcatc gagaaaatgc tcatccagga
     4861 tgccttgaag cggcacaggc acaatttcga cgcggtgctt caggagttgg agttgccaag
     4921 acgcaccctg tatcaccgca tgaaggaact gggagttgca gcgccgatcg ctgcgacggc
     4981 cggggtctaa taaaacgcag gtgtggttat gctactagag attaaagagg agaaatacta
     5041 gatgagtctt gatgaaaggt ttgaggatga tctggacgag gagcgggttc cgaatctggg
     5101 gatagttgcc gaaagtattt cgcaactggg tatcgacgtg ctgctatcgg gtgagaccgg
     5161 cacgggcaaa gacacgattg cccgacggat tcatgagatg tcaggccgca aagggcgcct
     5221 ggtggcgatg aattgcgcgg ccattccgga gtccctcgcc gagagcgagt tattcggcgt
     5281 ggtcagcggt gcctacaccg gcgctgatcg ctccagagtc ggttatgtcg aagcggcgca
     5341 gggcggcacg ctgtacctgg atgagatcga tagcatgccg ctgagcctgc aagccaaatt
     5401 gctgagggtg ctggaaaccc gagcgcttga acggctgggt tcgacgtcga cgatcaagct
     5461 ggatatctgc gtgatcgcct ccgcccaatg ctcgctggac gacgccgtcg agcgggggca
     5521 gtttcgtcgc gatctgtatt ttcgcctgaa cgtcctgaca ctcaagcttc ctccgctacg
     5581 taaccagtct gatcgcatag ttcccctgtt cacacgtttt acggccgccg ccgcgaggga
     5641 gctcggtgtt cccgttcccg atgtttgccc actgctgcac aaagtgctgc tgggccacga
     5701 ctggcccggc aatatccgtg agctcaaggc ggcagccaaa cgccatgtgc tgggtttccc
     5761 cttgctgggc gccgagccgc agggcgaaga gcacttggcc tgtgggctca aatcgcaatt
     5821 gcgagtgatc gaaaaagccc tgattcagga gtcgctcaag cgccacgaca attgtgtgga
     5881 ttcggtaagc ctggaactgg acgtgccacg ccgtacgctc tatcgacgca tcaaagaatt
     5941 gcagatctaa taatactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     6001 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     6061 caccttcggg tgggcctttc tgcgtttata tactagaggc cggattatgt ccgctgagtg
     6121 ggtcacggtc ccggatcagt tcccttgcga agctgaccga tgtttttgtg ccaaaagctg
     6181 ttgtggcaaa aaacggtttg cgcaaagttt tgtattacaa agaatttcac attttaaaat
     6241 atctttataa atcaatcagt tatttctatt ttcaagctgg cacggttatt gctatagggc
     6301 ttgtactact agagattaaa gaggagaaat actagatgac taaaaagaaa tatggattaa
     6361 aattatcaac agttcgaaag ttagaagatg agttgtgtga ttatcctaat tatcataagc
     6421 aactcgaaga tttaagaagt gaaataatga caccatggat tccaacagat acaaatatag
     6481 gcggggagtt tgtaccgtct aatacatcga aaacagaaat ggcagtaact aattatcttt
     6541 gtagtatacg aagaggtaaa atccttgagt ttaagagcgc tattgaacgt ataatcaaca
     6601 catcaagtag gaaagaacgc gaatttattc aagagtatta ttttaataaa aaggaattag
     6661 tgaaagtttg tgatgacata cacatttctg atagaactgc tcatagaatc aaaaggaaaa
     6721 tcatatccag attggcggaa gagttagggg aagagtgata ctagagccag gcatcaaata
     6781 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     6841 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacta
     6901 gagaattggc agtaaagtgg cagtttttga tacctaaaat gagatattat gatagtgtag
     6961 gatattgact atcttactgc gtttccctta tcgcaattag gaataaagga tctatgtggg
     7021 ttggctgatt atagccaatc cttttttaat tttaaaaagc gtatagcgcg agtactagag
     7081 attaaagagg agaaatacta gatgcgtaaa ggagaagaac ttttcactgg agttgtccca
     7141 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     7201 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     7261 cctgttccat ggccaacact tgtcactact ttcggttatg gtgttcaatg ctttgcgaga
     7321 tacccagatc atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     7381 caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag
     7441 tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat
     7501 ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt atacatcatg
     7561 gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat
     7621 ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     7681 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     7741 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     7801 gatgaactat acaaataata atactagagc caggcatcaa ataaaacgaa aggctcagtc
     7861 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     7921 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagtagcgg ccgctgca
//
