LOCUS       sequence-177384-        3322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 3 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term 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 3322)
  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 3322)
  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-STRNDHFR
FEATURES             Location/Qualifiers
     source          1..3322
                     /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             128..151
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             173..193
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(754..772)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      768..815
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        880..971
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1836..1875
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1906..2494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2395..2414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2648..2665
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2680..2819
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2745..2764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2905..2927
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2921..3112)
                     /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 ggcaagctgg agccacccgc agttcgagaa aggatctggt ggtggtggtg gtgaaaacct
      181 gtacttccag ggatccatca gtctgattgc ggcgctagcg gtagatcgcg ttatcggcat
      241 ggaaaacgcc atgccatgga acctgcctgc cgatctcgcc tggtttaaac gcaacacctt
      301 aaataaaccc gtgattatgg ggcgccatac ctgggaatca atcggtaggc ctttgcccgg
      361 ccgcaaaaat attatcctca gcagtcaacc cgggaccgat gatcgggtta cctgggttaa
      421 atcggtcgac gaagccatcg cggcgtgtgg tgacgtacca gaaatcatgg tgattggcgg
      481 cggacgcgtt tatgaacagt tcttgccaaa agcgcaaaaa ctgtatctga cgcatatcga
      541 tgcagaagtg gaaggcgaca cccattttcc ggattacgag ccggatgact gggaatcggt
      601 attcagcgaa ttccacgatg ctgatgcgca gaactcgcat agctattgtt tcgaaatcct
      661 cgagcgtcgt gccggctaat gtacaggtac cacgcgtgcg cgctgatccg gctgctaaca
      721 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
      781 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatcgggat
      841 tagcccgcct aatgagcggg ctttttttta attcccctat ttgtttattt ttctaaatac
      901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1021 tttgccttcc tgtttttgct cacccagaaa cgctcgtgaa agtaaaagac gcagaggacc
     1081 aattgggggc acgagtggga tacatagaac tggacttgaa tagcggtaaa atccttgaga
     1141 gttttcgccc tgaagagcgt tttccaatga tgagcacttt caaagttctg ctatgtggag
     1201 cagtattatc ccgtgtagat gcggggcaag agcaactcgg acgacgaata cactattcgc
     1261 agaatgactt ggttgaatac tccccagtga cagaaaagca ccttacggac ggaatgacgg
     1321 taagagaatt atgtagtgcc gccataacga tgagtgataa cactgcggcg aacttacttc
     1381 tgacaaccat cggtggaccg aaggaattaa ccgctttttt gcacaatatg ggagaccatg
     1441 taactcgcct tgaccgttgg gaaccagaac tgaatgaagc cataccaaac gacgagcgag
     1501 acaccacaat gcctgcggca atggcaacaa cattacgcaa actattaact ggcgaactac
     1561 ttactctggc ttcacggcaa caattaatag actggcttga agcggataaa gttgcaggac
     1621 cactactgcg ttcggcactt cctgctggct ggtttattgc tgataaatct ggggcaggag
     1681 agcgtggttc acggggtatc attgccgcac ttggaccaga tggtaagcct tcccgtatcg
     1741 tagttatcta cacgacgggt agtcaggcaa ctatggacga acgaaataga cagattgctg
     1801 aaataggggc ttcactgatt aagcattggt aaaccgatac aattaaaggc tccttttgga
     1861 gccttttttt ttggacggac cggtagaaaa gatcaaagga tcttcttgag atcctttttt
     1921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2041 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2521 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2581 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2641 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2701 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     2761 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     2821 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2881 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2941 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     3001 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     3061 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     3121 ccgtgtaagg gggatttctg ttcatggggt aatgataccg atgaaacgag agaggatgct
     3181 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     3241 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     3301 cgttaataca gatgtaggtg tt
//
