LOCUS       sequence-177380-        3280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 3 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 3280)
  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 3280)
  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-FLAGDHFR
FEATURES             Location/Qualifiers
     source          1..3280
                     /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"
     primer_bind     complement(712..730)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      726..773
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        838..929
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1794..1833
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1864..2452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2353..2372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2606..2623
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2638..2777
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2703..2722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2863..2885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2879..3070)
                     /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 atccatcagt ctgattgcgg cgctagcggt
      181 agatcgcgtt atcggcatgg aaaacgccat gccatggaac ctgcctgccg atctcgcctg
      241 gtttaaacgc aacaccttaa ataaacccgt gattatgggg cgccatacct gggaatcaat
      301 cggtaggcct ttgcccggcc gcaaaaatat tatcctcagc agtcaacccg ggaccgatga
      361 tcgggttacc tgggttaaat cggtcgacga agccatcgcg gcgtgtggtg acgtaccaga
      421 aatcatggtg attggcggcg gacgcgttta tgaacagttc ttgccaaaag cgcaaaaact
      481 gtatctgacg catatcgatg cagaagtgga aggcgacacc cattttccgg attacgagcc
      541 ggatgactgg gaatcggtat tcagcgaatt ccacgatgct gatgcgcaga actcgcatag
      601 ctattgtttc gaaatcctcg agcgtcgtgc cggctaatgt acaggtacca cgcgtgcgcg
      661 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      721 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      781 gaggaactat atcgggatta gcccgcctaa tgagcgggct tttttttaat tcccctattt
      841 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      901 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      961 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag
     1021 taaaagacgc agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata
     1081 gcggtaaaat ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca
     1141 aagttctgct atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac
     1201 gacgaataca ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc
     1261 ttacggacgg aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca
     1321 ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc
     1381 acaatatggg agaccatgta actcgccttg accgttggga accagaactg aatgaagcca
     1441 taccaaacga cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac
     1501 tattaactgg cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag
     1561 cggataaagt tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg
     1621 ataaatctgg ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg
     1681 gtaagccttc ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac
     1741 gaaatagaca gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa
     1801 ttaaaggctc cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc
     1861 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1921 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1981 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2041 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2101 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2161 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2221 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2281 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2341 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2401 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2461 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2521 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2581 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2641 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     2701 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     2761 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2821 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2881 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     2941 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3001 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3061 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catggggtaa tgataccgat
     3121 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3181 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3241 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
//
