LOCUS       sequence-177370-        3373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term cleavable double 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 3373)
  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 3373)
  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-STRHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3373
                     /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"
     CDS             173..190
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             218..238
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(805..823)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      819..866
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        931..1022
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1887..1926
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1957..2545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2446..2465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2699..2716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2731..2870
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2796..2815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2956..2978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2972..3163)
                     /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 aaaggatcta gccatcatca
      181 tcatcatcac agcagcggat ctggtggtgg tggtggtgaa aacctgtact tccagggatc
      241 catcagtctg attgcggcgc tagcggtaga tcgcgttatc ggcatggaaa acgccatgcc
      301 atggaacctg cctgccgatc tcgcctggtt taaacgcaac accttaaata aacccgtgat
      361 tatggggcgc catacctggg aatcaatcgg taggcctttg cccggccgca aaaatattat
      421 cctcagcagt caacccggga ccgatgatcg ggttacctgg gttaaatcgg tcgacgaagc
      481 catcgcggcg tgtggtgacg taccagaaat catggtgatt ggcggcggac gcgtttatga
      541 acagttcttg ccaaaagcgc aaaaactgta tctgacgcat atcgatgcag aagtggaagg
      601 cgacacccat tttccggatt acgagccgga tgactgggaa tcggtattca gcgaattcca
      661 cgatgctgat gcgcagaact cgcatagcta ttgtttcgaa atcctcgagc gtcgtgccgg
      721 ctaatgtaca aagcttggta ccacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa
      781 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      841 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatcggga ttagcccgcc
      901 taatgagcgg gctttttttt aattccccta tttgtttatt tttctaaata cattcaaata
      961 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1021 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1081 ctgtttttgc tcacccagaa acgctcgtga aagtaaaaga cgcagaggac caattggggg
     1141 cacgagtggg atacatagaa ctggacttga atagcggtaa aatccttgag agttttcgcc
     1201 ctgaagagcg ttttccaatg atgagcactt tcaaagttct gctatgtgga gcagtattat
     1261 cccgtgtaga tgcggggcaa gagcaactcg gacgacgaat acactattcg cagaatgact
     1321 tggttgaata ctccccagtg acagaaaagc accttacgga cggaatgacg gtaagagaat
     1381 tatgtagtgc cgccataacg atgagtgata acactgcggc gaacttactt ctgacaacca
     1441 tcggtggacc gaaggaatta accgcttttt tgcacaatat gggagaccat gtaactcgcc
     1501 ttgaccgttg ggaaccagaa ctgaatgaag ccataccaaa cgacgagcga gacaccacaa
     1561 tgcctgcggc aatggcaaca acattacgca aactattaac tggcgaacta cttactctgg
     1621 cttcacggca acaattaata gactggcttg aagcggataa agttgcagga ccactactgc
     1681 gttcggcact tcctgctggc tggtttattg ctgataaatc tggggcagga gagcgtggtt
     1741 cacggggtat cattgccgca cttggaccag atggtaagcc ttcccgtatc gtagttatct
     1801 acacgacggg tagtcaggca actatggacg aacgaaatag acagattgct gaaatagggg
     1861 cttcactgat taagcattgg taaaccgata caattaaagg ctccttttgg agcctttttt
     1921 tttggacgga ccggtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1981 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2041 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2101 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2161 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2221 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2281 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2341 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2401 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2461 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2521 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2581 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2641 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2701 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     2761 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2821 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     2881 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2941 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3001 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3061 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3121 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     3181 ggggatttct gttcatgggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     3241 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     3301 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     3361 agatgtaggt gtt
//
