LOCUS       sequence-64445-e         870 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 870)
  AUTHORS   Sathiamoorthy S, Shin JA
  TITLE     Boundaries of the origin of replication: creation of a 
            pET-28a-derived vector with p15A copy control allowing compatible 
            coexistence with pET vectors.
  JOURNAL   PLoS One. 2012;7(10):e47259. doi: 10.1371/journal.pone.0047259. Epub
            2012 Oct 22.
  PUBMED    23110063
REFERENCE   2  (bases 1 to 870)
  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: "PLoS One.";
            date: "2012"; volume: "7(10):e47259. doi"; pages: " 
            10.1371/journal.pone.0047259. Epub 2012 Oct 22"
COMMENT     Sequence Label: pSAM
FEATURES             Location/Qualifiers
     source          1..870
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(20..37)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(87..119)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(123..140)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(150..167)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             186..208
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    223..247
                     /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(247..266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(248..266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(350..369)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        575..652
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(672..691)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gttagcagcc ggaatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
       61 cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagccata
      121 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
      181 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
      241 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
      301 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
      361 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
      421 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
      481 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
      541 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
      601 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
      661 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
      721 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
      781 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
      841 gattggcgtt gccacctcag tctggcctgc
//
