LOCUS       sequence-177508-        3286 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 3286)
  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 3286)
  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-HISDHFR
FEATURES             Location/Qualifiers
     source          1..3286
                     /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             131..148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(717..735)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      731..778
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        843..934
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1799..1838
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1869..2457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2358..2377
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2611..2628
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2643..2782
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2708..2727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2868..2890
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2884..3075)
                     /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 gggcagcagc catcatcatc atcatcacag cagcggatcc atcagtctga ttgcggcgct
      181 agcggtagat cgcgttatcg gcatggaaaa cgccatgcca tggaacctgc ctgccgatct
      241 cgcctggttt aaacgcaaca ccttaaataa acccgtgatt atggggcgcc atacctggga
      301 atcaatcggt aggcctttgc ccggccgcaa aaatattatc ctcagcagtc aacccgggac
      361 cgatgatcgg gttacctggg ttaaatcggt cgacgaagcc atcgcggcgt gtggtgacgt
      421 accagaaatc atggtgattg gcggcggacg cgtttatgaa cagttcttgc caaaagcgca
      481 aaaactgtat ctgacgcata tcgatgcaga agtggaaggc gacacccatt ttccggatta
      541 cgagccggat gactgggaat cggtattcag cgaattccac gatgctgatg cgcagaactc
      601 gcatagctat tgtttcgaaa tcctcgagcg tcgtgccggc taatgtacgg taccacgcgt
      661 gcgcgctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      721 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      781 gaaaggagga actatatcgg gattagcccg cctaatgagc gggctttttt ttaattcccc
      841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctcgt
     1021 gaaagtaaaa gacgcagagg accaattggg ggcacgagtg ggatacatag aactggactt
     1081 gaatagcggt aaaatccttg agagttttcg ccctgaagag cgttttccaa tgatgagcac
     1141 tttcaaagtt ctgctatgtg gagcagtatt atcccgtgta gatgcggggc aagagcaact
     1201 cggacgacga atacactatt cgcagaatga cttggttgaa tactccccag tgacagaaaa
     1261 gcaccttacg gacggaatga cggtaagaga attatgtagt gccgccataa cgatgagtga
     1321 taacactgcg gcgaacttac ttctgacaac catcggtgga ccgaaggaat taaccgcttt
     1381 tttgcacaat atgggagacc atgtaactcg ccttgaccgt tgggaaccag aactgaatga
     1441 agccatacca aacgacgagc gagacaccac aatgcctgcg gcaatggcaa caacattacg
     1501 caaactatta actggcgaac tacttactct ggcttcacgg caacaattaa tagactggct
     1561 tgaagcggat aaagttgcag gaccactact gcgttcggca cttcctgctg gctggtttat
     1621 tgctgataaa tctggggcag gagagcgtgg ttcacggggt atcattgccg cacttggacc
     1681 agatggtaag ccttcccgta tcgtagttat ctacacgacg ggtagtcagg caactatgga
     1741 cgaacgaaat agacagattg ctgaaatagg ggcttcactg attaagcatt ggtaaaccga
     1801 tacaattaaa ggctcctttt ggagcctttt tttttggacg gaccggtaga aaagatcaaa
     1861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1981 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2221 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2281 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2341 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2401 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2461 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2521 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2581 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2641 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     2701 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     2761 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     2821 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2881 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     2941 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     3001 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     3061 ttcctgtttg gtcactgatg ccttccgtgt aagggggatt tctgttcatg gggtaatgat
     3121 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3181 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3241 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgtt
//
