LOCUS       sequence-248602-        4306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arsenic sensor used in the sensor cell array. pSB3K3 carrying 
            J105-30arsR-t-ParsR-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4306)
  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 4306)
  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: As5 (pBW414J105-arsR)
FEATURES             Location/Qualifiers
     source          1..4306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(140..955)
                     /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(140..955)
                     /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     863..882
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     947..966
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(1550..2094)
                     /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(1629..1648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2562..2580
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2584..2627)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2630..2651
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2652..2686
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     RBS             2735..2746
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3169..3240
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3256..3283
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3411..3422
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3429..4142
                     /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(3459..3487)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4082..4102
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4165..4236
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4252..4279
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4286..4306
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
       61 cactcaaagg cggtaatctc gagtcccgtc aagtcagcgt aatgctctgc cagtgttaca
      121 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
      181 tcatatcagg attatcgata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      241 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      301 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      361 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
      421 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      481 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      541 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
      601 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
      661 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
      721 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
      781 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
      841 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
      901 tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg ctcataacac
      961 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     1021 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     1081 cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg
     1141 caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg
     1201 gctccctcac tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac
     1261 cttcttcacg aggcagacct cagcgctagc ggagtgtata ctggcttact atgttggcac
     1321 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     1381 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     1441 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     1501 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     1561 tccgccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     1621 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     1681 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     1741 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     1801 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     1861 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     1921 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     1981 cagttacctc ggttcaaaga gttggtagct cagagaacct tcaaaaaacc gccctgcaag
     2041 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     2101 catcttatta aggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattccatg gtgccacctg
     2521 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2581 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc
     2641 cgcttctaga gtttacggct agctcagtcc taggtactat gctagctact agtagctagc
     2701 atagtaccta ggactgagct agctactaga gattaaagag gagaaatact agatgtcatt
     2761 tctgttaccc atccaattgt tcaaaattct tgctgatgaa acccgtctgg gcatcgtttt
     2821 actgctcagc gaactgggag agttatgcgt ctgcgatctc tgcactgctc tcgaccagtc
     2881 gcagcccaag atctcccgcc acctggcatt gctgcgtgaa agcgggctat tgctggaccg
     2941 caagcaaggt aagtgggttc attaccgctt atcaccgcat attccagcat gggcggcgaa
     3001 aattattgat gaggcctggc gatgtgaaca ggaaaaggtt caggcgattg tccgcaacct
     3061 ggctcgacaa aactgttccg gggacagtaa gaacatttgc agttaaaaat ttagctaaac
     3121 acacatgaat tttcagatgt gttttatccg ggtactagag ccaggcatca aataaaacga
     3181 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     3241 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat actagagcca
     3301 actcaaaatt cacacctatt accttcctct gcacttacac attcgttaag tcatatatgt
     3361 ttttgactta tccgcttcga agagagacac tacctgcaat actagagatt aaagaggaga
     3421 aatactagat gcgtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     3481 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     3541 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     3601 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
     3661 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     3721 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     3781 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     3841 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
     3901 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     3961 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     4021 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     4081 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     4141 aataataata ctagagccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     4201 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
     4261 tcgggtgggc ctttctgcgt ttatatacta gtagcggccg ctgcag
//
