LOCUS       sequence-311324-        2749 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Two-layered AHL-responsive circuit carrying an ECF11-based 
            amplifier.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2749)
  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 2749)
  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: pXW101Plux2-Pecf11-gfp
FEATURES             Location/Qualifiers
     source          1..2749
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        1..35
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             47..58
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      862..933
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      949..976
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1675..1746
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1762..1789
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1875..1886
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1893..2606
                     /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(1923..1951)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2546..2566
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2629..2700
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2716..2743
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tttacagcta gctcagtcct aggtattatg ctagctacta gagattaaag aggagaaata
       61 ctagatgaaa aacataaatg ccgacgacac atacagaata attaataaaa ttaaagcttg
      121 tagaagcaat aatgatatta atcaatgctt atctgatatg actaaaatgg tacattgtga
      181 atattattta ctcgcgatca tttatcctca ttctatggtt aaatctgata tttcaatcct
      241 agataattac cctaaaaaat ggaggcaata ttatgatgac gctaatttaa taaaatatga
      301 tcctatagta gattattcta actccaatca ttcaccaatt aattggaata tatttgaaaa
      361 caatgctgta aataaaaaat ctccaaatgt aattaaagaa gcgaaaacat caggtcttat
      421 cactgggttt agtttcccta ttcatacggc taacaatggc ttcggaatgc ttagttttgc
      481 acattcagaa aaagacaact atatagatag tttattttta catgcgtgta tgaacatacc
      541 attaattgtt ccttctctag ttgataatta tcgaaaaata aatatagcaa ataataaatc
      601 aaacaacgat ttaaccaaaa gagaaaaaga atgtttagcg tgggcatgcg aaggaaaaag
      661 ctcttgggat atttcaaaaa tattaggttg cagtgagcgt actgtcactt tccatttaac
      721 caatgcgcaa atgaaactca atacaacaaa ccgctgccaa agtatttcta aagcaatttt
      781 aacaggagca attgattgcc catactttaa aaattaataa cactgatagt gctagtgtag
      841 atcactacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      901 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
      961 ggtgggcctt tctgcgttta tatactagag acctgtagga tcgtacaggt ttacgcaaga
     1021 aaatggtttg ttactttcga ataaatacta gagtcacaca ggactactag atgatgagcg
     1081 atagtccgca gaaactgggt cgtaatgaat ggaatgcata tatggataaa gtgaaagcca
     1141 aagatcgtga agcctttgcc tttgtgtttc gtttttatgc accgaaactg aaacagttcg
     1201 cctataaaca tgttggcaat gaacaggttg caatggaaat ggttcaagaa accatggcaa
     1261 ccgtttggca gaaagcacat ctgtatgatg gtaaaaaaag cgcactgagc acctggattt
     1321 ataccattat tcgtaacctg tgctttgacc tgctgcgtaa acagaaaggt aaagaactgc
     1381 atattcacag cgacgatatt tggccgagcg aatattatcc gcctgatatg gttgatcatt
     1441 atagtccgga acaggatatg ctgaaagaac aggtggttaa atttctggat atcctgccga
     1501 aaaatcagcg tgatgttctg caagcagttt atctggaaga actgccgcat cagcaggttg
     1561 cagaactgtt tgatattccg ctgggcaccg ttaaaagccg tctgcgtctg gcagttgaaa
     1621 aactgcgtca tagcatgcat accgaacagc tgtaataata ctagagccag gcatcaaata
     1681 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1741 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacta
     1801 gaggcctcca caccgctcgt cacatcctgt gatccactct tcatcccgct acgtaacacc
     1861 tcttactaga gattaaagag gagaaatact agatgcgtaa aggagaagaa cttttcactg
     1921 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     1981 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     2041 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcggttat ggtgttcaat
     2101 gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg
     2161 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     2221 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     2281 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg
     2341 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca
     2401 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     2461 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     2521 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     2581 cacatggcat ggatgaacta tacaaataat aatactagag ccaggcatca aataaaacga
     2641 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2701 tactagagtc acactggctc accttcgggt gggcctttct gcgtttata
//
