LOCUS       sequence-177396-        3325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3325)
  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 3325)
  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: pST50Trc4-CBPDHFR
FEATURES             Location/Qualifiers
     source          1..3325
                     /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             83..105
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             119..196
                     /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(757..775)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      771..818
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        883..974
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1839..1878
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1909..2497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2398..2417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2651..2668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2683..2822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2748..2767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2908..2930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2924..3115)
                     /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 tccggaaata attttgttta actttaagaa ggagatatac atatgtctaa
      121 gcgacgatgg aaaaagaatt tcatagccgt ctcagcagcc aaccgcttta agaaaatctc
      181 atcctccggg gcacttctgg gatccatcag tctgattgcg gcgctagcgg tagatcgcgt
      241 tatcggcatg gaaaacgcca tgccatggaa cctgcctgcc gatctcgcct ggtttaaacg
      301 caacacctta aataaacccg tgattatggg gcgccatacc tgggaatcaa tcggtaggcc
      361 tttgcccggc cgcaaaaata ttatcctcag cagtcaaccc gggaccgatg atcgggttac
      421 ctgggttaaa tcggtcgacg aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt
      481 gattggcggc ggacgcgttt atgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac
      541 gcatatcgat gcagaagtgg aaggcgacac ccattttccg gattacgagc cggatgactg
      601 ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag aactcgcata gctattgttt
      661 cgaaatcctc gagcgtcgtg ccggctaatg tacaacgcgt gcgcgctgat ccggctgcta
      721 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
      781 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatcgg
      841 gattagcccg cctaatgagc gggctttttt ttaattcccc tatttgttta tttttctaaa
      901 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      961 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1021 cattttgcct tcctgttttt gctcacccag aaacgctcgt gaaagtaaaa gacgcagagg
     1081 accaattggg ggcacgagtg ggatacatag aactggactt gaatagcggt aaaatccttg
     1141 agagttttcg ccctgaagag cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg
     1201 gagcagtatt atcccgtgta gatgcggggc aagagcaact cggacgacga atacactatt
     1261 cgcagaatga cttggttgaa tactccccag tgacagaaaa gcaccttacg gacggaatga
     1321 cggtaagaga attatgtagt gccgccataa cgatgagtga taacactgcg gcgaacttac
     1381 ttctgacaac catcggtgga ccgaaggaat taaccgcttt tttgcacaat atgggagacc
     1441 atgtaactcg ccttgaccgt tgggaaccag aactgaatga agccatacca aacgacgagc
     1501 gagacaccac aatgcctgcg gcaatggcaa caacattacg caaactatta actggcgaac
     1561 tacttactct ggcttcacgg caacaattaa tagactggct tgaagcggat aaagttgcag
     1621 gaccactact gcgttcggca cttcctgctg gctggtttat tgctgataaa tctggggcag
     1681 gagagcgtgg ttcacggggt atcattgccg cacttggacc agatggtaag ccttcccgta
     1741 tcgtagttat ctacacgacg ggtagtcagg caactatgga cgaacgaaat agacagattg
     1801 ctgaaatagg ggcttcactg attaagcatt ggtaaaccga tacaattaaa ggctcctttt
     1861 ggagcctttt tttttggacg gaccggtaga aaagatcaaa ggatcttctt gagatccttt
     1921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2041 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2161 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2221 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2281 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2341 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2401 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2461 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2521 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2581 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2641 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2701 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2761 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2821 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2881 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2941 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3001 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3061 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3121 cctccgtgta agggggattt ctgttcatgg ggtaatgata ccgatgaaac gagagaggat
     3181 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3241 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     3301 cttcgttaat acagatgtag gtgtt
//
