LOCUS       sequence-177492-         719 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Position 2 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 719)
  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 719)
  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: pST50Trc2-CBPHISNDHFR
FEATURES             Location/Qualifiers
     source          1..719
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(412..432)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(460..477)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(490..567)
                     /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             581..603
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(658..677)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(659..677)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaataccgat tcccagtcat ccggctcgta atccggaaaa tgggtgtcgc cttccacttc
       61 tgcatcgata tgcgtcagat acagtttttg cgcttttggc aagaactgtt cataaacgcg
      121 tccgccgcca atcaccatga tttctggtac gtcaccacac gccgcgatgg cttcgtcgac
      181 cgatttaacc caggtaaccc gatcatcggt cccgggttga ctgctgagga taatattttt
      241 gcggccgggc aaaggcctac cgattgattc ccaggtatgg cgccccataa tcacgggttt
      301 atttaaggtg ttgcgtttaa accaggcgag atcggcaggc aggttccatg gcatggcgtt
      361 ttccatgccg ataacgcgat ctaccgctag cgccgcaatc agactgatgg atccctggaa
      421 gtacaggttt tcaccaccac caccaccaga tccgctgctg tgatgatgat gatgatggct
      481 agatcccaga agtgccccgg aggatgagat tttcttaaag cggttggctg ctgagacggc
      541 tatgaaattc tttttccatc gtcgcttaga catatgtata tctccttctt aaagttaaac
      601 aaaattattg aattccaatt ggagctctcc ggagccggag ggaaaccgtt gtggtctccc
      661 tatagtgagt cgtattaatt tcgcgggatc gatgcatggt gccaacacct acatctggt
//
