LOCUS       sequence-243995-        8103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arsenic sensor with two-layered amplifier cascade. pSB4A3 carrying 
            J117-30arsR-t-ParsR-30hrpR-30hrpS-t-PhrpLE-31ECF11(ASV)-t-Pecf11-30g
            fp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8103)
  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 8103)
  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: pXW117As(RS-E11)
FEATURES             Location/Qualifiers
     source          1..8103
                     /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_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             3364..3375
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3798..3869
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3885..3912
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             4040..4051
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4962..4973
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             5036..5047
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      5982..6053
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6069..6096
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      7009..7080
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7096..7123
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             7209..7220
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7227..7940
                     /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(7257..7285)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7880..7900
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      7963..8034
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8050..8077
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    join(8084..8103,1)
                     /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 tctagagttg acagctagct cagtcctagg gattgtgcta gctactagag
     3361 attaaagagg agaaatacta gatgtcattt ctgttaccca tccaattgtt caaaattctt
     3421 gctgatgaaa cccgtctggg catcgtttta ctgctcagcg aactgggaga gttatgcgtc
     3481 tgcgatctct gcactgctct cgaccagtcg cagcccaaga tctcccgcca cctggcattg
     3541 ctgcgtgaaa gcgggctatt gctggaccgc aagcaaggta agtgggttca ttaccgctta
     3601 tcaccgcata ttccagcatg ggcggcgaaa attattgatg aggcctggcg atgtgaacag
     3661 gaaaaggttc aggcgattgt ccgcaacctg gctcgacaaa actgttccgg ggacagtaag
     3721 aacatttgca gttaaaaatt tagctaaaca cacatgaatt ttcagatgtg ttttatccgg
     3781 gtactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     3841 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     3901 ggcctttctg cgtttatata ctagagccaa ctcaaaattc acacctatta ccttcctctg
     3961 cacttacaca ttcgttaagt catatatgtt tttgacttat ccgcttcgaa gagagacact
     4021 acctgcaata ctagagatta aagaggagaa atactagatg agtacaggca tcgataagga
     4081 cgtccgagag tgttggggcg taactgcatt atcagcgggt catcaaattg caatgaatag
     4141 cgcgtttctg gatatggact tgctgttgtg cggggaaacc ggcaccggca aggacacact
     4201 ggccaaccgc attcacgagt tgtccagcag gtcgggaccc tttgtgggca tgaactgcgc
     4261 cgccattccc gagtcgctgg cagagagcca gttattcggt gtggtcaacg gtgcattcac
     4321 cggcgtatgc cgggctcgcg agggctacat agaggcctcc agtggtggca ccttgtacct
     4381 ggatgaaatc gacagcatgc cgttgagcct gcaagccaaa ctgctgcgtg tgttggagag
     4441 tcgaggtatc gagcgtctgg gctcgaccga atttatcccg gtggatctgc ggatcattgc
     4501 ctcggcccag cggccactgg atgaactggt ggaacaagga cttttccgtc gcgacctgtt
     4561 ttttcggctc aacgtgctga cgcttcactt gccagccttg cgcaaacgtc gtgaacagat
     4621 cctgccattg ttcgaccagt tcacccaggg tatcgctgcc gagttcggac gtcccgctcc
     4681 tgcgctggac agcgggcgtg tgcagctgct gctcagccac gactggccgg gcaacatccg
     4741 cgaattgaag tctgcggcca agcgcttcgt actcggcttc cccttgctgg gcgccgaccc
     4801 tgtggaagcg cttgaccctg ccacggggct gcgcacgcaa atgcgcatca tcgagaaaat
     4861 gctcatccag gatgccttga agcggcacag gcacaatttc gacgcggtgc ttcaggagtt
     4921 ggagttgcca agacgcaccc tgtatcaccg catgaaggaa ctgggagttg cagcgccgat
     4981 cgctgcgacg gccggggtct aataaaacgc aggtgtggtt atgctactag agattaaaga
     5041 ggagaaatac tagatgagtc ttgatgaaag gtttgaggat gatctggacg aggagcgggt
     5101 tccgaatctg gggatagttg ccgaaagtat ttcgcaactg ggtatcgacg tgctgctatc
     5161 gggtgagacc ggcacgggca aagacacgat tgcccgacgg attcatgaga tgtcaggccg
     5221 caaagggcgc ctggtggcga tgaattgcgc ggccattccg gagtccctcg ccgagagcga
     5281 gttattcggc gtggtcagcg gtgcctacac cggcgctgat cgctccagag tcggttatgt
     5341 cgaagcggcg cagggcggca cgctgtacct ggatgagatc gatagcatgc cgctgagcct
     5401 gcaagccaaa ttgctgaggg tgctggaaac ccgagcgctt gaacggctgg gttcgacgtc
     5461 gacgatcaag ctggatatct gcgtgatcgc ctccgcccaa tgctcgctgg acgacgccgt
     5521 cgagcggggg cagtttcgtc gcgatctgta ttttcgcctg aacgtcctga cactcaagct
     5581 tcctccgcta cgtaaccagt ctgatcgcat agttcccctg ttcacacgtt ttacggccgc
     5641 cgccgcgagg gagctcggtg ttcccgttcc cgatgtttgc ccactgctgc acaaagtgct
     5701 gctgggccac gactggcccg gcaatatccg tgagctcaag gcggcagcca aacgccatgt
     5761 gctgggtttc cccttgctgg gcgccgagcc gcagggcgaa gagcacttgg cctgtgggct
     5821 caaatcgcaa ttgcgagtga tcgaaaaagc cctgattcag gagtcgctca agcgccacga
     5881 caattgtgtg gattcggtaa gcctggaact ggacgtgcca cgccgtacgc tctatcgacg
     5941 catcaaagaa ttgcagatct aataatacta gagccaggca tcaaataaaa cgaaaggctc
     6001 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     6061 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag gccggattat
     6121 gtccgctgag tgggtcacgg tcccggatca gttcccttgc gaagctgacc gatgtttttg
     6181 tgccaaaagc tgttgtggca aaaaacggtt tgcgcaaagt tttgtattac aaagaatttc
     6241 acattttaaa atatctttat aaatcaatca gttatttcta ttttcaagct ggcacggtta
     6301 ttgctatagg gcttgtacta ctagagtcac acaggaaacc tactagatga tgagcgatag
     6361 tccgcagaaa ctgggtcgta atgaatggaa tgcatatatg gataaagtga aagccaaaga
     6421 tcgtgaagcc tttgcctttg tgtttcgttt ttatgcaccg aaactgaaac agttcgccta
     6481 taaacatgtt ggcaatgaac aggttgcaat ggaaatggtt caagaaacca tggcaaccgt
     6541 ttggcagaaa gcacatctgt atgatggtaa aaaaagcgca ctgagcacct ggatttatac
     6601 cattattcgt aacctgtgct ttgacctgct gcgtaaacag aaaggtaaag aactgcatat
     6661 tcacagcgac gatatttggc cgagcgaata ttatccgcct gatatggttg atcattatag
     6721 tccggaacag gatatgctga aagaacaggt ggttaaattt ctggatatcc tgccgaaaaa
     6781 tcagcgtgat gttctgcaag cagtttatct ggaagaactg ccgcatcagc aggttgcaga
     6841 actgtttgat attccgctgg gcaccgttaa aagccgtctg cgtctggcag ttgaaaaact
     6901 gcgtcatagc atgcataccg aacagctgag gcctgctgca aacgacgaaa actacgctgc
     6961 atcagtttaa taaaacgcag gtgtggttat gctactagag ccaggcatca aataaaacga
     7021 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     7081 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat actagaggcc
     7141 tccacaccgc tcgtcacatc ctgtgatcca ctcttcatcc cgctacgtaa cacctcttac
     7201 tagagattaa agaggagaaa tactagatgc gtaaaggaga agaacttttc actggagttg
     7261 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
     7321 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
     7381 aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt caatgctttg
     7441 cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg cccgaaggtt
     7501 atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag
     7561 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag
     7621 aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca
     7681 tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg
     7741 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     7801 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     7861 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     7921 gcatggatga actatacaaa taataatact agagccaggc atcaaataaa acgaaaggct
     7981 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     8041 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggccgct
     8101 gca
//
