LOCUS       sequence-240627-        6614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amplified arsenic sensor with double ArsR binding sites. pSB3K3 
            carrying J109-30arsR-t-ParsR-ABS84-30hrpR-30hrpS-t-PhrpLE-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6614)
  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 6614)
  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: pXW109ABS84(RS)
FEATURES             Location/Qualifiers
     source          1..6614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..21
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(161..976)
                     /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(161..976)
                     /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     884..903
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     968..987
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(1571..2115)
                     /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(1650..1669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2583..2601
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2605..2648)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2651..2672
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2673..2707
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     RBS             2719..2730
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3153..3224
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3240..3267
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3458..3469
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4380..4391
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             4454..4465
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      5392..5463
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5479..5506
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             5740..5751
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5758..6471
                     /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(5788..5816)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6411..6431
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      6494..6565
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6581..6608
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tactagtagc ggccgctgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
       61 ctgcggcgag cggtatcagc tcactcaaag gcggtaatct cgagtcccgt caagtcagcg
      121 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
      181 tcaaatgaaa ctgcaattta ttcatatcag gattatcgat accatatttt tgaaaaagcc
      241 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
      301 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
      361 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
      421 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
      481 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
      541 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
      601 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
      661 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
      721 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
      781 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
      841 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
      901 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc
      961 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     1021 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     1081 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     1141 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     1201 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     1261 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat
     1321 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     1381 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     1441 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     1501 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     1561 caaagccgtt tttccatagg ctccgccccc tgacaagcat cacgaaatct gacgctcaaa
     1621 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc
     1681 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     1741 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     1801 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     1861 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     1921 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     1981 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     2041 ttcaaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     2101 ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaac
     2161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2281 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2341 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2401 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2461 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2521 tctattccat ggtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     2581 aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg
     2641 atgatttctg gaattcgcgg ccgcttctag agtttacagc tagctcagtc ctagggactg
     2701 tgctagctac tagagattaa agaggagaaa tactagatgt catttctgtt acccatccaa
     2761 ttgttcaaaa ttcttgctga tgaaacccgt ctgggcatcg ttttactgct cagcgaactg
     2821 ggagagttat gcgtctgcga tctctgcact gctctcgacc agtcgcagcc caagatctcc
     2881 cgccacctgg cattgctgcg tgaaagcggg ctattgctgg accgcaagca aggtaagtgg
     2941 gttcattacc gcttatcacc gcatattcca gcatgggcgg cgaaaattat tgatgaggcc
     3001 tggcgatgtg aacaggaaaa ggttcaggcg attgtccgca acctggctcg acaaaactgt
     3061 tccggggaca gtaagaacat ttgcagttaa aaatttagct aaacacacat gaattttcag
     3121 atgtgtttta tccgggtact agagccaggc atcaaataaa acgaaaggct cagtcgaaag
     3181 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     3241 gctcaccttc gggtgggcct ttctgcgttt atatactaga gccaactcaa aattcacacc
     3301 tattaccttc ctctgcactt acacattcgt taagtcatat atgtttttga cttatccgct
     3361 tcgaagagag acactacctg caactattac cttcctctgc acttacacat tcgttaagtc
     3421 atatatgttt ttgacttatc cgcttctact agagattaaa gaggagaaat actagatgag
     3481 tacaggcatc gataaggacg tccgagagtg ttggggcgta actgcattat cagcgggtca
     3541 tcaaattgca atgaatagcg cgtttctgga tatggacttg ctgttgtgcg gggaaaccgg
     3601 caccggcaag gacacactgg ccaaccgcat tcacgagttg tccagcaggt cgggaccctt
     3661 tgtgggcatg aactgcgccg ccattcccga gtcgctggca gagagccagt tattcggtgt
     3721 ggtcaacggt gcattcaccg gcgtatgccg ggctcgcgag ggctacatag aggcctccag
     3781 tggtggcacc ttgtacctgg atgaaatcga cagcatgccg ttgagcctgc aagccaaact
     3841 gctgcgtgtg ttggagagtc gaggtatcga gcgtctgggc tcgaccgaat ttatcccggt
     3901 ggatctgcgg atcattgcct cggcccagcg gccactggat gaactggtgg aacaaggact
     3961 tttccgtcgc gacctgtttt ttcggctcaa cgtgctgacg cttcacttgc cagccttgcg
     4021 caaacgtcgt gaacagatcc tgccattgtt cgaccagttc acccagggta tcgctgccga
     4081 gttcggacgt cccgctcctg cgctggacag cgggcgtgtg cagctgctgc tcagccacga
     4141 ctggccgggc aacatccgcg aattgaagtc tgcggccaag cgcttcgtac tcggcttccc
     4201 cttgctgggc gccgaccctg tggaagcgct tgaccctgcc acggggctgc gcacgcaaat
     4261 gcgcatcatc gagaaaatgc tcatccagga tgccttgaag cggcacaggc acaatttcga
     4321 cgcggtgctt caggagttgg agttgccaag acgcaccctg tatcaccgca tgaaggaact
     4381 gggagttgca gcgccgatcg ctgcgacggc cggggtctaa taaaacgcag gtgtggttat
     4441 gctactagag attaaagagg agaaatacta gatgagtctt gatgaaaggt ttgaggatga
     4501 tctggacgag gagcgggttc cgaatctggg gatagttgcc gaaagtattt cgcaactggg
     4561 tatcgacgtg ctgctatcgg gtgagaccgg cacgggcaaa gacacgattg cccgacggat
     4621 tcatgagatg tcaggccgca aagggcgcct ggtggcgatg aattgcgcgg ccattccgga
     4681 gtccctcgcc gagagcgagt tattcggcgt ggtcagcggt gcctacaccg gcgctgatcg
     4741 ctccagagtc ggttatgtcg aagcggcgca gggcggcacg ctgtacctgg atgagatcga
     4801 tagcatgccg ctgagcctgc aagccaaatt gctgagggtg ctggaaaccc gagcgcttga
     4861 acggctgggt tcgacgtcga cgatcaagct ggatatctgc gtgatcgcct ccgcccaatg
     4921 ctcgctggac gacgccgtcg agcgggggca gtttcgtcgc gatctgtatt ttcgcctgaa
     4981 cgtcctgaca ctcaagcttc ctccgctacg taaccagtct gatcgcatag ttcccctgtt
     5041 cacacgtttt acggccgccg ccgcgaggga gctcggtgtt cccgttcccg atgtttgccc
     5101 actgctgcac aaagtgctgc tgggccacga ctggcccggc aatatccgtg agctcaaggc
     5161 ggcagccaaa cgccatgtgc tgggtttccc cttgctgggc gccgagccgc agggcgaaga
     5221 gcacttggcc tgtgggctca aatcgcaatt gcgagtgatc gaaaaagccc tgattcagga
     5281 gtcgctcaag cgccacgaca attgtgtgga ttcggtaagc ctggaactgg acgtgccacg
     5341 ccgtacgctc tatcgacgca tcaaagaatt gcagatctaa taaccaggca tcaaataaaa
     5401 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     5461 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag
     5521 gccggattat gtccgctgag tgggtcacgg tcccggatca gttcccttgc gaagctgacc
     5581 gatgtttttg tgccaaaagc tgttgtggca aaaaacggtt tgcgcaaagt tttgtattac
     5641 aaagaatttc acattttaaa atatctttat aaatcaatca gttatttcta ttttcaagct
     5701 ggcacggtta ttgctatagg gcttgtacta ctagagatta aagaggagaa atactagatg
     5761 cgtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
     5821 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
     5881 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
     5941 actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcatat gaaacagcat
     6001 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa
     6061 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     6121 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     6181 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     6241 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
     6301 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
     6361 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
     6421 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa ataataatac
     6481 tagagccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     6541 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
     6601 tttctgcgtt tata
//
