LOCUS       sequence-240629-        6597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amplified arsenic sensor with double ArsR binding sites. pSB3K3 
            carrying J109-30arsR-t-ParsR-ABS67-30hrpR-30hrpS-t-PhrpLE-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6597)
  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 6597)
  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: pXW109ABS67(RS)
FEATURES             Location/Qualifiers
     source          1..6597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..56
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     RBS             68..79
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      502..573
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      589..616
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             790..801
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1712..1723
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             1786..1797
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2724..2795
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2811..2838
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3072..3083
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3090..3803
                     /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(3120..3148)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3743..3763
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3826..3897
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3913..3940
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3947..3967
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(4107..4922)
                     /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(4107..4922)
                     /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     4830..4849
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4914..4933
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5517..6061)
                     /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(5596..5615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6529..6547
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6551..6594)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(6597,1..21)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aattcgcggc cgcttctaga gtttacagct agctcagtcc tagggactgt gctagctact
       61 agagattaaa gaggagaaat actagatgtc atttctgtta cccatccaat tgttcaaaat
      121 tcttgctgat gaaacccgtc tgggcatcgt tttactgctc agcgaactgg gagagttatg
      181 cgtctgcgat ctctgcactg ctctcgacca gtcgcagccc aagatctccc gccacctggc
      241 attgctgcgt gaaagcgggc tattgctgga ccgcaagcaa ggtaagtggg ttcattaccg
      301 cttatcaccg catattccag catgggcggc gaaaattatt gatgaggcct ggcgatgtga
      361 acaggaaaag gttcaggcga ttgtccgcaa cctggctcga caaaactgtt ccggggacag
      421 taagaacatt tgcagttaaa aatttagcta aacacacatg aattttcaga tgtgttttat
      481 ccgggtacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      541 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
      601 ggtgggcctt tctgcgttta tatactagag ccaactcaaa attcacacct attaccttcc
      661 tctgcactta cacattcgtt aagtcatata tgtttttgac ttatccgctt cgaagagaga
      721 cactacctgc aaacttacac attcgttaag tcatatatgt ttttgactta tccgcttcta
      781 ctagagatta aagaggagaa atactagatg agtacaggca tcgataagga cgtccgagag
      841 tgttggggcg taactgcatt atcagcgggt catcaaattg caatgaatag cgcgtttctg
      901 gatatggact tgctgttgtg cggggaaacc ggcaccggca aggacacact ggccaaccgc
      961 attcacgagt tgtccagcag gtcgggaccc tttgtgggca tgaactgcgc cgccattccc
     1021 gagtcgctgg cagagagcca gttattcggt gtggtcaacg gtgcattcac cggcgtatgc
     1081 cgggctcgcg agggctacat agaggcctcc agtggtggca ccttgtacct ggatgaaatc
     1141 gacagcatgc cgttgagcct gcaagccaaa ctgctgcgtg tgttggagag tcgaggtatc
     1201 gagcgtctgg gctcgaccga atttatcccg gtggatctgc ggatcattgc ctcggcccag
     1261 cggccactgg atgaactggt ggaacaagga cttttccgtc gcgacctgtt ttttcggctc
     1321 aacgtgctga cgcttcactt gccagccttg cgcaaacgtc gtgaacagat cctgccattg
     1381 ttcgaccagt tcacccaggg tatcgctgcc gagttcggac gtcccgctcc tgcgctggac
     1441 agcgggcgtg tgcagctgct gctcagccac gactggccgg gcaacatccg cgaattgaag
     1501 tctgcggcca agcgcttcgt actcggcttc cccttgctgg gcgccgaccc tgtggaagcg
     1561 cttgaccctg ccacggggct gcgcacgcaa atgcgcatca tcgagaaaat gctcatccag
     1621 gatgccttga agcggcacag gcacaatttc gacgcggtgc ttcaggagtt ggagttgcca
     1681 agacgcaccc tgtatcaccg catgaaggaa ctgggagttg cagcgccgat cgctgcgacg
     1741 gccggggtct aataaaacgc aggtgtggtt atgctactag agattaaaga ggagaaatac
     1801 tagatgagtc ttgatgaaag gtttgaggat gatctggacg aggagcgggt tccgaatctg
     1861 gggatagttg ccgaaagtat ttcgcaactg ggtatcgacg tgctgctatc gggtgagacc
     1921 ggcacgggca aagacacgat tgcccgacgg attcatgaga tgtcaggccg caaagggcgc
     1981 ctggtggcga tgaattgcgc ggccattccg gagtccctcg ccgagagcga gttattcggc
     2041 gtggtcagcg gtgcctacac cggcgctgat cgctccagag tcggttatgt cgaagcggcg
     2101 cagggcggca cgctgtacct ggatgagatc gatagcatgc cgctgagcct gcaagccaaa
     2161 ttgctgaggg tgctggaaac ccgagcgctt gaacggctgg gttcgacgtc gacgatcaag
     2221 ctggatatct gcgtgatcgc ctccgcccaa tgctcgctgg acgacgccgt cgagcggggg
     2281 cagtttcgtc gcgatctgta ttttcgcctg aacgtcctga cactcaagct tcctccgcta
     2341 cgtaaccagt ctgatcgcat agttcccctg ttcacacgtt ttacggccgc cgccgcgagg
     2401 gagctcggtg ttcccgttcc cgatgtttgc ccactgctgc acaaagtgct gctgggccac
     2461 gactggcccg gcaatatccg tgagctcaag gcggcagcca aacgccatgt gctgggtttc
     2521 cccttgctgg gcgccgagcc gcagggcgaa gagcacttgg cctgtgggct caaatcgcaa
     2581 ttgcgagtga tcgaaaaagc cctgattcag gagtcgctca agcgccacga caattgtgtg
     2641 gattcggtaa gcctggaact ggacgtgcca cgccgtacgc tctatcgacg catcaaagaa
     2701 ttgcagatct aataaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2761 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     2821 cgggtgggcc tttctgcgtt tatatactag aggccggatt atgtccgctg agtgggtcac
     2881 ggtcccggat cagttccctt gcgaagctga ccgatgtttt tgtgccaaaa gctgttgtgg
     2941 caaaaaacgg tttgcgcaaa gttttgtatt acaaagaatt tcacatttta aaatatcttt
     3001 ataaatcaat cagttatttc tattttcaag ctggcacggt tattgctata gggcttgtac
     3061 tactagagat taaagaggag aaatactaga tgcgtaaagg agaagaactt ttcactggag
     3121 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     3181 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     3241 gaaaactacc tgttccatgg ccaacacttg tcactacttt cggttatggt gttcaatgct
     3301 ttgcgagata cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     3361 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     3421 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     3481 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     3541 acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca
     3601 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     3661 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     3721 ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac
     3781 atggcatgga tgaactatac aaataataat actagagcca ggcatcaaat aaaacgaaag
     3841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     3901 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agtagcggcc
     3961 gctgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4021 atcagctcac tcaaaggcgg taatctcgag tcccgtcaag tcagcgtaat gctctgccag
     4081 tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc
     4141 aatttattca tatcaggatt atcgatacca tatttttgaa aaagccgttt ctgtaatgaa
     4201 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     4261 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     4321 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt
     4381 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     4441 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     4501 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     4561 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     4621 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     4681 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     4741 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     4801 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     4861 gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg aatatggctc
     4921 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     4981 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata
     5041 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     5101 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
     5161 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca
     5221 gcaacacctt cttcacgagg cagacctcag cgctagcgga gtgtatactg gcttactatg
     5281 ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc
     5341 ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct
     5401 acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct
     5461 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
     5521 cataggctcc gccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     5581 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     5641 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     5701 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     5761 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5821 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     5881 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     5941 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttca aaaaaccgcc
     6001 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     6061 agaagatcat cttattaagg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6121 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6181 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6241 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6301 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6361 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6421 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttccatggtg
     6481 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6541 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctgg
//
