LOCUS       sequence-177402-        3310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3310)
  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 3310)
  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-STRNDHFR
FEATURES             Location/Qualifiers
     source          1..3310
                     /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             122..145
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             167..187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(742..760)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      756..803
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        868..959
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1824..1863
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1894..2482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2383..2402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2636..2653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2668..2807
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2733..2752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2893..2915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2909..3100)
                     /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 atatggcaag
      121 ctggagccac ccgcagttcg agaaaggatc tggtggtggt ggtggtgaaa acctgtactt
      181 ccagggatcc atcagtctga ttgcggcgct agcggtagat cgcgttatcg gcatggaaaa
      241 cgccatgcca tggaacctgc ctgccgatct cgcctggttt aaacgcaaca ccttaaataa
      301 acccgtgatt atggggcgcc atacctggga atcaatcggt aggcctttgc ccggccgcaa
      361 aaatattatc ctcagcagtc aacccgggac cgatgatcgg gttacctggg ttaaatcggt
      421 cgacgaagcc atcgcggcgt gtggtgacgt accagaaatc atggtgattg gcggcggacg
      481 cgtttatgaa cagttcttgc caaaagcgca aaaactgtat ctgacgcata tcgatgcaga
      541 agtggaaggc gacacccatt ttccggatta cgagccggat gactgggaat cggtattcag
      601 cgaattccac gatgctgatg cgcagaactc gcatagctat tgtttcgaaa tcctcgagcg
      661 tcgtgccggc taatgtacaa cgcgtgcgcg ctgatccggc tgctaacaaa gcccgaaagg
      721 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      781 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atcgggatta gcccgcctaa
      841 tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt
      901 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      961 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1021 tttttgctca cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa ttgggggcac
     1081 gagtgggata catagaactg gacttgaata gcggtaaaat ccttgagagt tttcgccctg
     1141 aagagcgttt tccaatgatg agcactttca aagttctgct atgtggagca gtattatccc
     1201 gtgtagatgc ggggcaagag caactcggac gacgaataca ctattcgcag aatgacttgg
     1261 ttgaatactc cccagtgaca gaaaagcacc ttacggacgg aatgacggta agagaattat
     1321 gtagtgccgc cataacgatg agtgataaca ctgcggcgaa cttacttctg acaaccatcg
     1381 gtggaccgaa ggaattaacc gcttttttgc acaatatggg agaccatgta actcgccttg
     1441 accgttggga accagaactg aatgaagcca taccaaacga cgagcgagac accacaatgc
     1501 ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg cgaactactt actctggctt
     1561 cacggcaaca attaatagac tggcttgaag cggataaagt tgcaggacca ctactgcgtt
     1621 cggcacttcc tgctggctgg tttattgctg ataaatctgg ggcaggagag cgtggttcac
     1681 ggggtatcat tgccgcactt ggaccagatg gtaagccttc ccgtatcgta gttatctaca
     1741 cgacgggtag tcaggcaact atggacgaac gaaatagaca gattgctgaa ataggggctt
     1801 cactgattaa gcattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt
     1861 ggacggaccg gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2041 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2581 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2641 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     2701 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2761 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     2821 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2881 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2941 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3001 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3061 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     3121 gatttctgtt catggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3181 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3241 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3301 tgtaggtgtt
//
