LOCUS       sequence-177397-        3271 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Position 4 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 3271)
  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 3271)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-STRDHFR
FEATURES             Location/Qualifiers
     source          1..3271
                     /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"
     primer_bind     complement(703..721)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      717..764
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        829..920
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1785..1824
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1855..2443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2344..2363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2597..2614
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2629..2768
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2694..2713)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2854..2876
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2870..3061)
                     /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 catcagtctg attgcggcgc tagcggtaga
      181 tcgcgttatc ggcatggaaa acgccatgcc atggaacctg cctgccgatc tcgcctggtt
      241 taaacgcaac accttaaata aacccgtgat tatggggcgc catacctggg aatcaatcgg
      301 taggcctttg cccggccgca aaaatattat cctcagcagt caacccggga ccgatgatcg
      361 ggttacctgg gttaaatcgg tcgacgaagc catcgcggcg tgtggtgacg taccagaaat
      421 catggtgatt ggcggcggac gcgtttatga acagttcttg ccaaaagcgc aaaaactgta
      481 tctgacgcat atcgatgcag aagtggaagg cgacacccat tttccggatt acgagccgga
      541 tgactgggaa tcggtattca gcgaattcca cgatgctgat gcgcagaact cgcatagcta
      601 ttgtttcgaa atcctcgagc gtcgtgccgg ctaatgtaca acgcgtgcgc gctgatccgg
      661 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
      721 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
      781 tatcgggatt agcccgccta atgagcgggc ttttttttaa ttcccctatt tgtttatttt
      841 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      901 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      961 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg
     1021 cagaggacca attgggggca cgagtgggat acatagaact ggacttgaat agcggtaaaa
     1081 tccttgagag ttttcgccct gaagagcgtt ttccaatgat gagcactttc aaagttctgc
     1141 tatgtggagc agtattatcc cgtgtagatg cggggcaaga gcaactcgga cgacgaatac
     1201 actattcgca gaatgacttg gttgaatact ccccagtgac agaaaagcac cttacggacg
     1261 gaatgacggt aagagaatta tgtagtgccg ccataacgat gagtgataac actgcggcga
     1321 acttacttct gacaaccatc ggtggaccga aggaattaac cgcttttttg cacaatatgg
     1381 gagaccatgt aactcgcctt gaccgttggg aaccagaact gaatgaagcc ataccaaacg
     1441 acgagcgaga caccacaatg cctgcggcaa tggcaacaac attacgcaaa ctattaactg
     1501 gcgaactact tactctggct tcacggcaac aattaataga ctggcttgaa gcggataaag
     1561 ttgcaggacc actactgcgt tcggcacttc ctgctggctg gtttattgct gataaatctg
     1621 gggcaggaga gcgtggttca cggggtatca ttgccgcact tggaccagat ggtaagcctt
     1681 cccgtatcgt agttatctac acgacgggta gtcaggcaac tatggacgaa cgaaatagac
     1741 agattgctga aataggggct tcactgatta agcattggta aaccgataca attaaaggct
     1801 ccttttggag cctttttttt tggacggacc ggtagaaaag atcaaaggat cttcttgaga
     1861 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1921 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1981 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2041 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2101 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2161 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2221 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2281 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2341 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2401 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2461 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2521 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2581 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     2641 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     2701 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2761 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2821 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2881 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     2941 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     3001 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     3061 ctgatgcctc cgtgtaaggg ggatttctgt tcatggggta atgataccga tgaaacgaga
     3121 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     3181 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     3241 ccagcgcttc gttaatacag atgtaggtgt t
//
