LOCUS       sequence-94442-e         978 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial expression of PDB 4NW3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 978)
  TITLE     SGC_Toronto_Q4_2013_Q1_2014
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 978)
  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: 
            "Unpublished"
COMMENT     Sequence Label: JMC044-B12
FEATURES             Location/Qualifiers
     source          1..978
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     64..83
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        64..82
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    83..107
                     /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             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             154..171
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             184..204
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             408..425
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(478..496)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      492..539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      576..956
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(663..682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     873..894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
       61 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
      121 ttttgtttaa ctttaagaag gagatatacc atgcatcatc atcatcatca cagcagcggc
      181 agagaaaact tgtatttcca gggcaagaaa ggacgtcgat cgaggcggtg tgggcagtgt
      241 cccggctgcc aggtgcctga ggactgtggt gtttgtacta attgcttaga taagcccaag
      301 tttggtggtc gcaatataaa gaagcagtgc tgcaagatga gaaaatgtca gaatctacaa
      361 tggatgcctt ccaaagcctg atgacgaagc ttgcggccgc actcgagcac caccaccacc
      421 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      481 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      541 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
      601 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      661 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      721 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      781 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      841 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      901 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
      961 ttggttaaaa aatgagct
//
