LOCUS       sequence-140676-         531 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  T7 promoter, RBS, His6-tag and thrombin cleave site - derived from 
            pET15b (5' CTAT/3' CATA) fusion).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 531)
  AUTHORS   Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont
            PS
  TITLE     EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2016 May 2.
  PUBMED    27096716
REFERENCE   2  (bases 1 to 531)
  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: "ACS Synth 
            Biol. 2016 May 2."
COMMENT     Sequence Label: pBP-T7_RBS-His6-Thrombin
FEATURES             Location/Qualifiers
     source          1..531
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(21..64)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    67..86
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     126..145
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        126..144
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    145..169
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             184..206
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             225..242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             252..269
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      304..361
                     /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(385..402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
       61 tttctggaat tcgcggccgc ttctagaatc tagagacgca tatgggtctc actatcccgc
      121 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
      181 aattttgttt aactttaaga aggagatata ccatgggcag cagccatcat catcatcatc
      241 acagcagcgg cctggtgccg cgcggcagcc ataagagacc gcatgccgtc tcattagctg
      301 cagtccggca aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
      361 attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      421 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      481 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc t
//
