LOCUS       sequence-91761-e         999 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 999)
  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 999)
  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..999
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     60..79
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        60..78
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    79..103
                     /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             118..140
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(201..220)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             393..404
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             478..495
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(548..566)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      562..609
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(651..668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(865..884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
       61 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      121 gtttaacttt aagaaggaga tataccatgg ttgaacaaga tggattgcac gcaggttctc
      181 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
      241 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
      301 acctgtccgg tgccctgaat gaacctggtg gctctggcgc tcagcttaag aaagagctgc
      361 aggcaaacaa gaaagagctg gctcagctga agtgggaact gcaggcactg aagaaagaac
      421 tggctcagta aggatccgaa ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac
      481 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      541 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      601 ggttttttgc tgaaaggagg aactatatcc gacaggatat attccgcttc ctcgctcact
      661 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
      721 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
      781 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
      841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
      901 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
      961 tgtctcattc cacgcctgac actcagttcc gggtaggca
//
