LOCUS       sequence-141125-         851 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 851)
  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 851)
  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..851
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     4..21
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(45..102)
                     /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."
     CDS             complement(137..154)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(164..181)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             200..222
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    237..261
                     /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)."
     primer_bind     complement(261..280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(262..280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    320..339
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     terminator      342..385
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(389..407)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(635..654)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgcagcgagt cagtgagcga ggaagcctgc ataacgcgaa gtaatctttt cggttttaaa
       61 gaaaaagggc agggtggtga caccttgccc ttttttgccg gactgcagct aatgagacgg
      121 catgcggtct cttatggctg ccgcgcggca ccaggccgct gctgtgatga tgatgatgat
      181 ggctgctgcc catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
      241 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atagtgagac
      301 ccatatgcgt ctctagattc tagaagcggc cgcgaattcc agaaatcatc cttagcgaaa
      361 gctaaggatt ttttttatct gaaattctgc ctcgtgatac gcctattttt ataggttaat
      421 gtcatgataa taatggtttc ttagacgtca ggtggcactc gagttgatcg ggcacgtaag
      481 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
      541 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
      601 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
      661 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
      721 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
      781 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
      841 ttacaccgtt t
//
