LOCUS       sequence-177349-        3361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 1 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 3361)
  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 3361)
  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: pST50Trc1-FLAGNDHFR
FEATURES             Location/Qualifiers
     source          1..3361
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             194..214
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(793..811)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      807..854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        919..1010
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1875..1914
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1945..2533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2434..2453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2687..2704
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2719..2858
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2784..2803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2944..2966
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2960..3151)
                     /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 tccaattgga attcgctagc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatgggaga ctacaaggac gatgatgaca agggatctgg
      181 tggtggtggt ggtgaaaacc tgtacttcca gggatccatc agtctgattg cggcgctagc
      241 ggtagatcgc gttatcggca tggaaaacgc catgccatgg aacctgcctg ccgatctcgc
      301 ctggtttaaa cgcaacacct taaataaacc cgtgattatg gggcgccata cctgggaatc
      361 aatcggtagg cctttgcccg gccgcaaaaa tattatcctc agcagtcaac ccgggaccga
      421 tgatcgggtt acctgggtta aatcggtcga cgaagccatc gcggcgtgtg gtgacgtacc
      481 agaaatcatg gtgattggcg gcggacgcgt ttatgaacag ttcttgccaa aagcgcaaaa
      541 actgtatctg acgcatatcg atgcagaagt ggaaggcgac acccattttc cggattacga
      601 gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc agaactcgca
      661 tagctattgt ttcgaaatcc tcgagcgtcg tgccggctaa tgtacaagat ccagatctaa
      721 gcttggtacc acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
      781 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      841 tgaggggttt tttgctgaaa ggaggaacta tatcgggatt agcccgccta atgagcgggc
      901 ttttttttaa ttcccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1021 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1081 acccagaaac gctcgtgaaa gtaaaagacg cagaggacca attgggggca cgagtgggat
     1141 acatagaact ggacttgaat agcggtaaaa tccttgagag ttttcgccct gaagagcgtt
     1201 ttccaatgat gagcactttc aaagttctgc tatgtggagc agtattatcc cgtgtagatg
     1261 cggggcaaga gcaactcgga cgacgaatac actattcgca gaatgacttg gttgaatact
     1321 ccccagtgac agaaaagcac cttacggacg gaatgacggt aagagaatta tgtagtgccg
     1381 ccataacgat gagtgataac actgcggcga acttacttct gacaaccatc ggtggaccga
     1441 aggaattaac cgcttttttg cacaatatgg gagaccatgt aactcgcctt gaccgttggg
     1501 aaccagaact gaatgaagcc ataccaaacg acgagcgaga caccacaatg cctgcggcaa
     1561 tggcaacaac attacgcaaa ctattaactg gcgaactact tactctggct tcacggcaac
     1621 aattaataga ctggcttgaa gcggataaag ttgcaggacc actactgcgt tcggcacttc
     1681 ctgctggctg gtttattgct gataaatctg gggcaggaga gcgtggttca cggggtatca
     1741 ttgccgcact tggaccagat ggtaagcctt cccgtatcgt agttatctac acgacgggta
     1801 gtcaggcaac tatggacgaa cgaaatagac agattgctga aataggggct tcactgatta
     1861 agcattggta aaccgataca attaaaggct ccttttggag cctttttttt tggacggacc
     1921 ggtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2641 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2701 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2761 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2821 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2881 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2941 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3001 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3061 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3121 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     3181 tcatggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     3241 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3301 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     3361 t
//
