LOCUS       sequence-65247-e         804 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 804)
  AUTHORS   Shin J, Noireaux V
  TITLE     Efficient cell-free expression with the endogenous E. Coli RNA 
            polymerase and sigma factor 70.
  JOURNAL   J Biol Eng. 2010 Jun 24;4:8.
  PUBMED    20576148
REFERENCE   2  (bases 1 to 804)
  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: "J Biol 
            Eng."; date: "2010-06-24"; volume: "4"; pages: "8"
COMMENT     Sequence Label: pBEST-p15A-PLtetO1-tetR-T500
FEATURES             Location/Qualifiers
     source          1..804
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             57..79
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    103..121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    128..146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(417..435)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        503..607
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat ggtatatctc
       61 cttcttaaag ttaaacaaaa ttattgctag ctgtgctcag tatctctatc actgataggg
      121 atgtcaatct ctatcactga tagggagcat gcccagcgga acagcacaca catgtcgcgg
      181 tggttacgtc cgtcacgttc acgcatcagg cggatatcgt tagcccaccc agcaaaattc
      241 ggttttctgg ctgatggtgc gatagtcttc accatgtcaa acatccactc tgcggcggtc
      301 aggtcttctg ctgtccccca cttgctggcg ctctgaattg cagcatccgg tttcaccaca
      361 gaaaggtcgt tttctggctg gtcagaggat tcgccagaat tcttgaagac gaaagggcct
      421 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      481 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      541 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      601 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      661 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      721 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      781 tcgccccgaa gaacgttttc caat
//
