LOCUS       sequence-177342-        3379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 1 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term non-cleavable single tag; contains DHFR gene in BamHI-NgoMIV 
            cassette as positive control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3379)
  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 3379)
  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: pST50Trc1-CBPDHFR
FEATURES             Location/Qualifiers
     source          1..3379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..226
                     /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"
     primer_bind     complement(811..829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      825..872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        937..1028
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1893..1932
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1963..2551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2452..2471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2705..2722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2737..2876
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2802..2821)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2962..2984
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2978..3169)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggcaccatgc atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
       61 tccctccggc tccggagagc tccaattgga attcgctagc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatgtctaa gcgacgatgg aaaaagaatt tcatagccgt
      181 ctcagcagcc aaccgcttta agaaaatctc atcctccggg gcacttctgg gatccatcag
      241 tctgattgcg gcgctagcgg tagatcgcgt tatcggcatg gaaaacgcca tgccatggaa
      301 cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg tgattatggg
      361 gcgccatacc tgggaatcaa tcggtaggcc tttgcccggc cgcaaaaata ttatcctcag
      421 cagtcaaccc gggaccgatg atcgggttac ctgggttaaa tcggtcgacg aagccatcgc
      481 ggcgtgtggt gacgtaccag aaatcatggt gattggcggc ggacgcgttt atgaacagtt
      541 cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgat gcagaagtgg aaggcgacac
      601 ccattttccg gattacgagc cggatgactg ggaatcggta ttcagcgaat tccacgatgc
      661 tgatgcgcag aactcgcata gctattgttt cgaaatcctc gagcgtcgtg ccggctaatg
      721 tacaagatcc agatctaagc ttggtaccac gcgtgcgcgc tgatccggct gctaacaaag
      781 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      841 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tcgggattag
      901 cccgcctaat gagcgggctt ttttttaatt cccctatttg tttatttttc taaatacatt
      961 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1021 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1081 gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca gaggaccaat
     1141 tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc cttgagagtt
     1201 ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta tgtggagcag
     1261 tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac tattcgcaga
     1321 atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga atgacggtaa
     1381 gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac ttacttctga
     1441 caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga gaccatgtaa
     1501 ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac gagcgagaca
     1561 ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc gaactactta
     1621 ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt gcaggaccac
     1681 tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg gcaggagagc
     1741 gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc cgtatcgtag
     1801 ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag attgctgaaa
     1861 taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc ttttggagcc
     1921 tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2101 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2641 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2701 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2761 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     2821 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2881 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2941 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3001 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3061 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3121 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3181 tgtaaggggg atttctgttc atggggtaat gataccgatg aaacgagaga ggatgctcac
     3241 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     3301 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3361 taatacagat gtaggtgtt
//
