LOCUS       sequence-311323-        2403 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Two-layered mercury responsive circuit carrying an ECF11-based 
            amplifier.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2403)
  AUTHORS   Wan X, Pinto F, Yu L, Wang B
  TITLE     Synthetic protein-binding DNA sponge as a tool to tune gene 
            expression and mitigate protein toxicity.
  JOURNAL   Nat Commun. 2020 Nov 24;11(1):5961. doi: 10.1038/s41467-020-19552-9.
  PUBMED    33235249
REFERENCE   2  (bases 1 to 2403)
  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"; journalName: "Nat 
            Commun."; date: "2020-11-24"; volume: "11(1):5961. doi"; pages: " 
            10.1038/s41467-020-19552-9"
COMMENT     Sequence Label: pXW109PmerT-Pecf11-gfp
FEATURES             Location/Qualifiers
     source          1..2403
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        1..35
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     terminator      514..585
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      601..628
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1329..1400
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1416..1443
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1529..1540
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1547..2260
                     /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(1577..1605)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2200..2220
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2283..2354
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2370..2397
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tttacagcta gctcagtcct agggactgtg ctagctacta gagtcacaca ggaaagtact
       61 agatggaaaa taatttggaa aacctgacca ttggcgtttt tgccaaggcg gccggggtca
      121 acgtggagac aatccgcttc tatcagcgca agggcctgtt gcgggaaccg gacaagcctt
      181 acggcagcat ccgccgctat ggggaggcgg acgtggttcg ggtgaaattc gtgaaatcgg
      241 cacagcggct ggggttcagt ctggacgaga ttgccgagct gttgcggctc gacgatggca
      301 cccactgcga ggaggccagc agcctggccg aacacaagct caaggacgtg cgcgagaaga
      361 tggccgactt ggcgcgcatg gaaaccgtgc tgtctgaact cgtgtgcgcc tgccatgcac
      421 gaaaggggaa tgtttcctgc ccgttgatcg cgtcactaca gggcgaagca ggcctggcaa
      481 ggtcagctat gccttagtac tagagccagg catcaaataa aacgaaaggc tcagtcgaaa
      541 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
      601 ggctcacctt cgggtgggcc tttctgcgtt tatatggatc cgttccatat cgcttgacta
      661 cgtacatgag tacggaagta aggttacgct atccaatcct actagagtca cacaggacta
      721 ctagatgatg agcgatagtc cgcagaaact gggtcgtaat gaatggaatg catatatgga
      781 taaagtgaaa gccaaagatc gtgaagcctt tgcctttgtg tttcgttttt atgcaccgaa
      841 actgaaacag ttcgcctata aacatgttgg caatgaacag gttgcaatgg aaatggttca
      901 agaaaccatg gcaaccgttt ggcagaaagc acatctgtat gatggtaaaa aaagcgcact
      961 gagcacctgg atttatacca ttattcgtaa cctgtgcttt gacctgctgc gtaaacagaa
     1021 aggtaaagaa ctgcatattc acagcgacga tatttggccg agcgaatatt atccgcctga
     1081 tatggttgat cattatagtc cggaacagga tatgctgaaa gaacaggtgg ttaaatttct
     1141 ggatatcctg ccgaaaaatc agcgtgatgt tctgcaagca gtttatctgg aagaactgcc
     1201 gcatcagcag gttgcagaac tgtttgatat tccgctgggc accgttaaaa gccgtctgcg
     1261 tctggcagtt gaaaaactgc gtcatagcat gcataccgaa cagctgtaat aatactagag
     1321 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1381 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     1441 gcgtttatat actagaggcc tccacaccgc tcgtcacatc ctgtgatcca ctcttcatcc
     1501 cgctacgtaa cacctcttac tagagattaa agaggagaaa tactagatgc gtaaaggaga
     1561 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
     1621 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
     1681 atttatttgc actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg
     1741 ttatggtgtt caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa
     1801 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
     1861 ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
     1921 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
     1981 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt
     2041 caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa
     2101 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc
     2161 tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac
     2221 agctgctggg attacacatg gcatggatga actatacaaa taataatact agagccaggc
     2281 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2341 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     2401 ata
//
