LOCUS       sequence-241974-        6592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amplified arsenic sensor with double ArsR binding sites. pSB3K3 
            carrying J109-30arsR-t-ParsR-ABS62-30hrpR-30hrpS-t-PhrpLE-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  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 6592)
  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: pXW109ABS62(RS)
FEATURES             Location/Qualifiers
     source          1..6592
                     /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             785..796
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1707..1718
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             1781..1792
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2719..2790
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2806..2833
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3067..3078
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3085..3798
                     /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(3115..3143)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3738..3758
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3821..3892
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3908..3935
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3942..3962
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(4102..4917)
                     /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(4102..4917)
                     /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     4825..4844
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4909..4928
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5512..6056)
                     /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(5591..5610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6524..6542
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6546..6589)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(6592,1..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 ataagtcata tatgtttttg acttatccgc ttctactaga
      781 gattaaagag gagaaatact agatgagtac aggcatcgat aaggacgtcc gagagtgttg
      841 gggcgtaact gcattatcag cgggtcatca aattgcaatg aatagcgcgt ttctggatat
      901 ggacttgctg ttgtgcgggg aaaccggcac cggcaaggac acactggcca accgcattca
      961 cgagttgtcc agcaggtcgg gaccctttgt gggcatgaac tgcgccgcca ttcccgagtc
     1021 gctggcagag agccagttat tcggtgtggt caacggtgca ttcaccggcg tatgccgggc
     1081 tcgcgagggc tacatagagg cctccagtgg tggcaccttg tacctggatg aaatcgacag
     1141 catgccgttg agcctgcaag ccaaactgct gcgtgtgttg gagagtcgag gtatcgagcg
     1201 tctgggctcg accgaattta tcccggtgga tctgcggatc attgcctcgg cccagcggcc
     1261 actggatgaa ctggtggaac aaggactttt ccgtcgcgac ctgttttttc ggctcaacgt
     1321 gctgacgctt cacttgccag ccttgcgcaa acgtcgtgaa cagatcctgc cattgttcga
     1381 ccagttcacc cagggtatcg ctgccgagtt cggacgtccc gctcctgcgc tggacagcgg
     1441 gcgtgtgcag ctgctgctca gccacgactg gccgggcaac atccgcgaat tgaagtctgc
     1501 ggccaagcgc ttcgtactcg gcttcccctt gctgggcgcc gaccctgtgg aagcgcttga
     1561 ccctgccacg gggctgcgca cgcaaatgcg catcatcgag aaaatgctca tccaggatgc
     1621 cttgaagcgg cacaggcaca atttcgacgc ggtgcttcag gagttggagt tgccaagacg
     1681 caccctgtat caccgcatga aggaactggg agttgcagcg ccgatcgctg cgacggccgg
     1741 ggtctaataa aacgcaggtg tggttatgct actagagatt aaagaggaga aatactagat
     1801 gagtcttgat gaaaggtttg aggatgatct ggacgaggag cgggttccga atctggggat
     1861 agttgccgaa agtatttcgc aactgggtat cgacgtgctg ctatcgggtg agaccggcac
     1921 gggcaaagac acgattgccc gacggattca tgagatgtca ggccgcaaag ggcgcctggt
     1981 ggcgatgaat tgcgcggcca ttccggagtc cctcgccgag agcgagttat tcggcgtggt
     2041 cagcggtgcc tacaccggcg ctgatcgctc cagagtcggt tatgtcgaag cggcgcaggg
     2101 cggcacgctg tacctggatg agatcgatag catgccgctg agcctgcaag ccaaattgct
     2161 gagggtgctg gaaacccgag cgcttgaacg gctgggttcg acgtcgacga tcaagctgga
     2221 tatctgcgtg atcgcctccg cccaatgctc gctggacgac gccgtcgagc gggggcagtt
     2281 tcgtcgcgat ctgtattttc gcctgaacgt cctgacactc aagcttcctc cgctacgtaa
     2341 ccagtctgat cgcatagttc ccctgttcac acgttttacg gccgccgccg cgagggagct
     2401 cggtgttccc gttcccgatg tttgcccact gctgcacaaa gtgctgctgg gccacgactg
     2461 gcccggcaat atccgtgagc tcaaggcggc agccaaacgc catgtgctgg gtttcccctt
     2521 gctgggcgcc gagccgcagg gcgaagagca cttggcctgt gggctcaaat cgcaattgcg
     2581 agtgatcgaa aaagccctga ttcaggagtc gctcaagcgc cacgacaatt gtgtggattc
     2641 ggtaagcctg gaactggacg tgccacgccg tacgctctat cgacgcatca aagaattgca
     2701 gatctaataa ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2761 tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     2821 gggcctttct gcgtttatat actagaggcc ggattatgtc cgctgagtgg gtcacggtcc
     2881 cggatcagtt cccttgcgaa gctgaccgat gtttttgtgc caaaagctgt tgtggcaaaa
     2941 aacggtttgc gcaaagtttt gtattacaaa gaatttcaca ttttaaaata tctttataaa
     3001 tcaatcagtt atttctattt tcaagctggc acggttattg ctatagggct tgtactacta
     3061 gagattaaag aggagaaata ctagatgcgt aaaggagaag aacttttcac tggagttgtc
     3121 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     3181 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     3241 ctacctgttc catggccaac acttgtcact actttcggtt atggtgttca atgctttgcg
     3301 agatacccag atcatatgaa acagcatgac tttttcaaga gtgccatgcc cgaaggttat
     3361 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     3421 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
     3481 gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc
     3541 atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca caacattgaa
     3601 gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     3661 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     3721 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     3781 atggatgaac tatacaaata ataatactag agccaggcat caaataaaac gaaaggctca
     3841 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     3901 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagtag cggccgctgc
     3961 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4021 ctcactcaaa ggcggtaatc tcgagtcccg tcaagtcagc gtaatgctct gccagtgtta
     4081 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     4141 attcatatca ggattatcga taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     4201 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     4261 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     4321 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
     4381 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     4441 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4501 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     4561 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     4621 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     4681 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     4741 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     4801 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     4861 catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac
     4921 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     4981 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     5041 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     5101 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     5161 tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac
     5221 accttcttca cgaggcagac ctcagcgcta gcggagtgta tactggctta ctatgttggc
     5281 actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc
     5341 gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct
     5401 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
     5461 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     5521 gctccgcccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     5581 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
     5641 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
     5701 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     5761 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     5821 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     5881 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     5941 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcaaaaaa ccgccctgca
     6001 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     6061 atcatcttat taaggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattcca tggtgccacc
     6481 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6541 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct gg
//
