LOCUS       sequence-177361-        3301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 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 3301)
  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 3301)
  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: pST50Trc2-STRDHFR
FEATURES             Location/Qualifiers
     source          1..3301
                     /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             101..123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             140..163
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(733..751)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      747..794
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        859..950
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1815..1854
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1885..2473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2374..2393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2627..2644
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2659..2798
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2724..2743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2884..2906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2900..3091)
                     /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 attcaataat tttgtttaac tttaagaagg
      121 agatatacat atggcaagct ggagccaccc gcagttcgag aaaggatcca tcagtctgat
      181 tgcggcgcta gcggtagatc gcgttatcgg catggaaaac gccatgccat ggaacctgcc
      241 tgccgatctc gcctggttta aacgcaacac cttaaataaa cccgtgatta tggggcgcca
      301 tacctgggaa tcaatcggta ggcctttgcc cggccgcaaa aatattatcc tcagcagtca
      361 acccgggacc gatgatcggg ttacctgggt taaatcggtc gacgaagcca tcgcggcgtg
      421 tggtgacgta ccagaaatca tggtgattgg cggcggacgc gtttatgaac agttcttgcc
      481 aaaagcgcaa aaactgtatc tgacgcatat cgatgcagaa gtggaaggcg acacccattt
      541 tccggattac gagccggatg actgggaatc ggtattcagc gaattccacg atgctgatgc
      601 gcagaactcg catagctatt gtttcgaaat cctcgagcgt cgtgccggct aatgtacaaa
      661 gcttggtacc acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
      721 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      781 tgaggggttt tttgctgaaa ggaggaacta tatcgggatt agcccgccta atgagcgggc
      841 ttttttttaa ttcccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      901 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      961 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1021 acccagaaac gctcgtgaaa gtaaaagacg cagaggacca attgggggca cgagtgggat
     1081 acatagaact ggacttgaat agcggtaaaa tccttgagag ttttcgccct gaagagcgtt
     1141 ttccaatgat gagcactttc aaagttctgc tatgtggagc agtattatcc cgtgtagatg
     1201 cggggcaaga gcaactcgga cgacgaatac actattcgca gaatgacttg gttgaatact
     1261 ccccagtgac agaaaagcac cttacggacg gaatgacggt aagagaatta tgtagtgccg
     1321 ccataacgat gagtgataac actgcggcga acttacttct gacaaccatc ggtggaccga
     1381 aggaattaac cgcttttttg cacaatatgg gagaccatgt aactcgcctt gaccgttggg
     1441 aaccagaact gaatgaagcc ataccaaacg acgagcgaga caccacaatg cctgcggcaa
     1501 tggcaacaac attacgcaaa ctattaactg gcgaactact tactctggct tcacggcaac
     1561 aattaataga ctggcttgaa gcggataaag ttgcaggacc actactgcgt tcggcacttc
     1621 ctgctggctg gtttattgct gataaatctg gggcaggaga gcgtggttca cggggtatca
     1681 ttgccgcact tggaccagat ggtaagcctt cccgtatcgt agttatctac acgacgggta
     1741 gtcaggcaac tatggacgaa cgaaatagac agattgctga aataggggct tcactgatta
     1801 agcattggta aaccgataca attaaaggct ccttttggag cctttttttt tggacggacc
     1861 ggtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1921 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1981 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2041 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2101 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2161 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2221 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2281 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2341 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2401 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2461 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2521 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2581 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2641 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2701 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2761 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2821 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2881 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2941 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3001 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3061 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     3121 tcatggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     3181 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3241 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     3301 t
//
