LOCUS       sequence-177524-         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Position 3 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term cleavable double tag; contains DHFR gene in BamHI-NgoMIV 
            cassette as positive control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 886)
  AUTHORS   Tan S, Kern RC, Selleck W
  TITLE     The pST44 polycistronic expression system for producing protein 
            complexes in Escherichia coli.
  JOURNAL   Protein Expr Purif. 2005 Apr;40(2):385-95.
  PUBMED    15766881
REFERENCE   2  (bases 1 to 886)
  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: "Protein 
            Expr Purif."; date: "2005-04"; volume: "40(2)"; pages: "385-95"
COMMENT     Sequence Label: pST50Trc3-CBPHISNDHFR
FEATURES             Location/Qualifiers
     source          1..886
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(526..546)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(574..591)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(604..681)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     RBS             695..717
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(760..779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(761..779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttagcagccg gatcagcgcg cacgcgtggt acctgtacat tagccggcac gacgctcgag
       61 gatttcgaaa caatagctat gcgagttctg cgcatcagca tcgtggaatt cgctgaatac
      121 cgattcccag tcatccggct cgtaatccgg aaaatgggtg tcgccttcca cttctgcatc
      181 gatatgcgtc agatacagtt tttgcgcttt tggcaagaac tgttcataaa cgcgtccgcc
      241 gccaatcacc atgatttctg gtacgtcacc acacgccgcg atggcttcgt cgaccgattt
      301 aacccaggta acccgatcat cggtcccggg ttgactgctg aggataatat ttttgcggcc
      361 gggcaaaggc ctaccgattg attcccaggt atggcgcccc ataatcacgg gtttatttaa
      421 ggtgttgcgt ttaaaccagg cgagatcggc aggcaggttc catggcatgg cgttttccat
      481 gccgataacg cgatctaccg ctagcgccgc aatcagactg atggatccct ggaagtacag
      541 gttttcacca ccaccaccac cagatccgct gctgtgatga tgatgatgat ggctagatcc
      601 cagaagtgcc ccggaggatg agattttctt aaagcggttg gctgctgaga cggctatgaa
      661 attctttttc catcgtcgct tagacatatg tatatctcct tcttaaagtt aaacaaaatt
      721 attgagctct ccggagccgg agggaaaccg ttgtggtctc cctatagtga gtcgtattaa
      781 tttcgcggga tcgatgcatg gtgccaacac ctacatctgt attaacgaag cgctggcatt
      841 gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccag
//
