LOCUS       sequence-177389-        3352 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3352)
  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 3352)
  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-FLAGHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3352
                     /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             89..111
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             125..148
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             158..175
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             203..223
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(784..802)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      798..845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        910..1001
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1866..1905
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1936..2524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2425..2444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2678..2695
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2710..2849
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2775..2794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2935..2957
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2951..3142)
                     /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 tcaataattt tgtttaactt taagaaggag atatacatat
      121 gggagactac aaggacgatg atgacaaggg atctagccat catcatcatc atcacagcag
      181 cggatctggt ggtggtggtg gtgaaaacct gtacttccag ggatccatca gtctgattgc
      241 ggcgctagcg gtagatcgcg ttatcggcat ggaaaacgcc atgccatgga acctgcctgc
      301 cgatctcgcc tggtttaaac gcaacacctt aaataaaccc gtgattatgg ggcgccatac
      361 ctgggaatca atcggtaggc ctttgcccgg ccgcaaaaat attatcctca gcagtcaacc
      421 cgggaccgat gatcgggtta cctgggttaa atcggtcgac gaagccatcg cggcgtgtgg
      481 tgacgtacca gaaatcatgg tgattggcgg cggacgcgtt tatgaacagt tcttgccaaa
      541 agcgcaaaaa ctgtatctga cgcatatcga tgcagaagtg gaaggcgaca cccattttcc
      601 ggattacgag ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca
      661 gaactcgcat agctattgtt tcgaaatcct cgagcgtcgt gccggctaat gtacaggtac
      721 cacgcgtgcg cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      781 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      841 ttttgctgaa aggaggaact atatcgggat tagcccgcct aatgagcggg ctttttttta
      901 attcccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      961 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1021 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1081 cgctcgtgaa agtaaaagac gcagaggacc aattgggggc acgagtggga tacatagaac
     1141 tggacttgaa tagcggtaaa atccttgaga gttttcgccc tgaagagcgt tttccaatga
     1201 tgagcacttt caaagttctg ctatgtggag cagtattatc ccgtgtagat gcggggcaag
     1261 agcaactcgg acgacgaata cactattcgc agaatgactt ggttgaatac tccccagtga
     1321 cagaaaagca ccttacggac ggaatgacgg taagagaatt atgtagtgcc gccataacga
     1381 tgagtgataa cactgcggcg aacttacttc tgacaaccat cggtggaccg aaggaattaa
     1441 ccgctttttt gcacaatatg ggagaccatg taactcgcct tgaccgttgg gaaccagaac
     1501 tgaatgaagc cataccaaac gacgagcgag acaccacaat gcctgcggca atggcaacaa
     1561 cattacgcaa actattaact ggcgaactac ttactctggc ttcacggcaa caattaatag
     1621 actggcttga agcggataaa gttgcaggac cactactgcg ttcggcactt cctgctggct
     1681 ggtttattgc tgataaatct ggggcaggag agcgtggttc acggggtatc attgccgcac
     1741 ttggaccaga tggtaagcct tcccgtatcg tagttatcta cacgacgggt agtcaggcaa
     1801 ctatggacga acgaaataga cagattgctg aaataggggc ttcactgatt aagcattggt
     1861 aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacggac cggtagaaaa
     1921 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1981 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2041 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2101 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2161 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2221 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2281 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2341 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2401 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2461 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2521 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2581 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2641 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2701 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     2761 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     2821 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     2881 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2941 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3001 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3061 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3121 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggt
     3181 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3241 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3301 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg tt
//
