LOCUS       sequence-177554-        3340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3340)
  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 3340)
  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-FLAGHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3340
                     /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..142
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             197..217
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(772..790)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      786..833
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        898..989
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1854..1893
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1924..2512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2413..2432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2666..2683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2698..2837
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2763..2782)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2923..2945
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2939..3130)
                     /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 atatgggaga
      121 ctacaaggac gatgatgaca agggatctag ccatcatcat catcatcaca gcagcggatc
      181 tggtggtggt ggtggtgaaa acctgtactt ccagggatcc atcagtctga ttgcggcgct
      241 agcggtagat cgcgttatcg gcatggaaaa cgccatgcca tggaacctgc ctgccgatct
      301 cgcctggttt aaacgcaaca ccttaaataa acccgtgatt atggggcgcc atacctggga
      361 atcaatcggt aggcctttgc ccggccgcaa aaatattatc ctcagcagtc aacccgggac
      421 cgatgatcgg gttacctggg ttaaatcggt cgacgaagcc atcgcggcgt gtggtgacgt
      481 accagaaatc atggtgattg gcggcggacg cgtttatgaa cagttcttgc caaaagcgca
      541 aaaactgtat ctgacgcata tcgatgcaga agtggaaggc gacacccatt ttccggatta
      601 cgagccggat gactgggaat cggtattcag cgaattccac gatgctgatg cgcagaactc
      661 gcatagctat tgtttcgaaa tcctcgagcg tcgtgccggc taatgtacaa cgcgtgcgcg
      721 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      781 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      841 gaggaactat atcgggatta gcccgcctaa tgagcgggct tttttttaat tcccctattt
      901 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      961 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1021 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag
     1081 taaaagacgc agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata
     1141 gcggtaaaat ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca
     1201 aagttctgct atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac
     1261 gacgaataca ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc
     1321 ttacggacgg aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca
     1381 ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc
     1441 acaatatggg agaccatgta actcgccttg accgttggga accagaactg aatgaagcca
     1501 taccaaacga cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac
     1561 tattaactgg cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag
     1621 cggataaagt tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg
     1681 ataaatctgg ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg
     1741 gtaagccttc ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac
     1801 gaaatagaca gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa
     1861 ttaaaggctc cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc
     1921 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1981 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2041 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2101 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2161 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2221 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2281 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2341 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2401 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2461 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2521 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2581 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2641 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2701 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     2761 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     2821 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2881 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2941 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3001 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3061 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3121 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catggggtaa tgataccgat
     3181 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3241 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3301 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
//
