LOCUS       sequence-177400-        3313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3313)
  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 3313)
  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: pST50Trc4-HISNDHFR
FEATURES             Location/Qualifiers
     source          1..3313
                     /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             125..142
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             170..190
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(745..763)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      759..806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        871..962
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1827..1866
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1897..2485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2386..2405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2639..2656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2671..2810
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2736..2755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2896..2918
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2912..3103)
                     /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 atatgggcag
      121 cagccatcat catcatcatc acagcagcgg atctggtggt ggtggtggtg aaaacctgta
      181 cttccaggga tccatcagtc tgattgcggc gctagcggta gatcgcgtta tcggcatgga
      241 aaacgccatg ccatggaacc tgcctgccga tctcgcctgg tttaaacgca acaccttaaa
      301 taaacccgtg attatggggc gccatacctg ggaatcaatc ggtaggcctt tgcccggccg
      361 caaaaatatt atcctcagca gtcaacccgg gaccgatgat cgggttacct gggttaaatc
      421 ggtcgacgaa gccatcgcgg cgtgtggtga cgtaccagaa atcatggtga ttggcggcgg
      481 acgcgtttat gaacagttct tgccaaaagc gcaaaaactg tatctgacgc atatcgatgc
      541 agaagtggaa ggcgacaccc attttccgga ttacgagccg gatgactggg aatcggtatt
      601 cagcgaattc cacgatgctg atgcgcagaa ctcgcatagc tattgtttcg aaatcctcga
      661 gcgtcgtgcc ggctaatgta caacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa
      721 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      781 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatcggga ttagcccgcc
      841 taatgagcgg gctttttttt aattccccta tttgtttatt tttctaaata cattcaaata
      901 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      961 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1021 ctgtttttgc tcacccagaa acgctcgtga aagtaaaaga cgcagaggac caattggggg
     1081 cacgagtggg atacatagaa ctggacttga atagcggtaa aatccttgag agttttcgcc
     1141 ctgaagagcg ttttccaatg atgagcactt tcaaagttct gctatgtgga gcagtattat
     1201 cccgtgtaga tgcggggcaa gagcaactcg gacgacgaat acactattcg cagaatgact
     1261 tggttgaata ctccccagtg acagaaaagc accttacgga cggaatgacg gtaagagaat
     1321 tatgtagtgc cgccataacg atgagtgata acactgcggc gaacttactt ctgacaacca
     1381 tcggtggacc gaaggaatta accgcttttt tgcacaatat gggagaccat gtaactcgcc
     1441 ttgaccgttg ggaaccagaa ctgaatgaag ccataccaaa cgacgagcga gacaccacaa
     1501 tgcctgcggc aatggcaaca acattacgca aactattaac tggcgaacta cttactctgg
     1561 cttcacggca acaattaata gactggcttg aagcggataa agttgcagga ccactactgc
     1621 gttcggcact tcctgctggc tggtttattg ctgataaatc tggggcagga gagcgtggtt
     1681 cacggggtat cattgccgca cttggaccag atggtaagcc ttcccgtatc gtagttatct
     1741 acacgacggg tagtcaggca actatggacg aacgaaatag acagattgct gaaatagggg
     1801 cttcactgat taagcattgg taaaccgata caattaaagg ctccttttgg agcctttttt
     1861 tttggacgga ccggtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1921 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1981 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2041 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2101 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2161 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2221 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2281 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2341 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2401 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2461 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2521 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2581 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2641 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     2701 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2761 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     2821 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2881 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2941 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3001 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3061 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     3121 ggggatttct gttcatgggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     3181 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     3241 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     3301 agatgtaggt gtt
//
