LOCUS       sequence-147508-         809 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Post cloned 4-piece from Isothermal/Golden Gate assembly (GGA), 
            using sequences from 2 (PltetO1-UTR1), 3 (tetR+linker from primer 
            extension), 21 (linker from primer extension + deGFP), and 105 
            (vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 809)
  AUTHORS   Sun ZZ, Yeung E, Hayes CA, Noireaux V, Murray RM
  TITLE     Linear DNA for rapid prototyping of synthetic biological circuits in
            an Escherichia coli based TX-TL cell-free system.
  JOURNAL   ACS Synth Biol. 2014 Jun 20;3(6):387-97. doi: 10.1021/sb400131a. 
            Epub 2013 Dec 4.
  PUBMED    24303785
REFERENCE   2  (bases 1 to 809)
  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: "10.1021/sb400131a";
            journalName: "ACS Synth Biol"; date: "2014-06-20- 20"; volume: "3"; 
            issue: "6"; pages: "387-97"
COMMENT     Sequence Label: pBEST-Pl-tetO1-tetR-linker-deGFP-T500 (109p)
FEATURES             Location/Qualifiers
     source          1..809
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    42..60
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    67..85
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             109..131
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             140..760
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(213..232)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             764..784
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 gtaaccaccg cgacatgtgt gtgctgttcc gctgggcatg ctccctatca gtgatagaga
       61 ttgacatccc tatcagtgat agagatactg agcacagcta gcaataattt tgtttaactt
      121 taagaaggag atataccaca tgtctagatt agataaaagt aaagtgatta acagcgcatt
      181 agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct
      241 aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc
      301 cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag
      361 ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg
      421 cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga
      481 aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact
      541 cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc
      601 taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac gacaagctat
      661 cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat
      721 atgcggatta gaaaaacaac ttaaatgtga aagtgggtct ggtgaaaacc tgtacttcca
      781 gtctggtggt gctatggagc ttttcactg
//
