LOCUS       sequence-91765-e         981 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Produces one half (alpha) of aminoglycoside phosphotransferase 
            (3′)-IIa when in the presence of T7 RNA Polymerase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 981)
  AUTHORS   Schmidt CM, Shis DL, Nguyen-Huu TD, Bennett MR
  TITLE     Stable maintenance of multiple plasmids in E. coli using a single 
            selective marker.
  JOURNAL   ACS Synth Biol. 2012 Oct 19;1(10):445-50. doi: 10.1021/sb3000589. 
            Epub 2012 Sep 4.
  PUBMED    23656183
REFERENCE   2  (bases 1 to 981)
  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"; doi: "10.1021/sb3000589";
            journalName: "ACS Synth Biol"; date: "2012-10-19- 19"; volume: "1"; 
            issue: "10"; pages: "445-50"
COMMENT     Sequence Label: pCS4
FEATURES             Location/Qualifiers
     source          1..981
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(31..48)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(122..133)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     306..325
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     RBS             386..408
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    423..447
                     /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(447..466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(448..466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(550..569)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        775..852
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(872..891)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctttcgggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct cgagtgcggc
       61 cgcaagcttg tcgacggagc tcgaattcgg atccttactg agccagttct ttcttcagtg
      121 cctgcagttc ccacttcagc tgagccagct ctttcttgtt tgcctgcagc tctttcttaa
      181 gctgagcgcc agagccacca ggttcattca gggcaccgga caggtcggtc ttgacaaaaa
      241 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
      301 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
      361 atccatcttg ttcaaccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga
      421 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
      481 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
      541 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
      601 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
      661 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
      721 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
      781 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
      841 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
      901 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
      961 aaagtggaag cggcgatggc g
//
