LOCUS       sequence-177364-        3343 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Position 2 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 3343)
  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 3343)
  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: pST50Trc2-HISNDHFR
FEATURES             Location/Qualifiers
     source          1..3343
                     /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             143..160
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(775..793)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      789..836
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        901..992
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1857..1896
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1927..2515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2416..2435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2669..2686
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2701..2840
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2766..2785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2926..2948
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2942..3133)
                     /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 atgggcagca gccatcatca tcatcatcac agcagcggat ctggtggtgg
      181 tggtggtgaa aacctgtact tccagggatc catcagtctg attgcggcgc tagcggtaga
      241 tcgcgttatc ggcatggaaa acgccatgcc atggaacctg cctgccgatc tcgcctggtt
      301 taaacgcaac accttaaata aacccgtgat tatggggcgc catacctggg aatcaatcgg
      361 taggcctttg cccggccgca aaaatattat cctcagcagt caacccggga ccgatgatcg
      421 ggttacctgg gttaaatcgg tcgacgaagc catcgcggcg tgtggtgacg taccagaaat
      481 catggtgatt ggcggcggac gcgtttatga acagttcttg ccaaaagcgc aaaaactgta
      541 tctgacgcat atcgatgcag aagtggaagg cgacacccat tttccggatt acgagccgga
      601 tgactgggaa tcggtattca gcgaattcca cgatgctgat gcgcagaact cgcatagcta
      661 ttgtttcgaa atcctcgagc gtcgtgccgg ctaatgtaca aagcttggta ccacgcgtgc
      721 gcgctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      781 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      841 aaggaggaac tatatcggga ttagcccgcc taatgagcgg gctttttttt aattccccta
      901 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
      961 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1021 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctcgtga
     1081 aagtaaaaga cgcagaggac caattggggg cacgagtggg atacatagaa ctggacttga
     1141 atagcggtaa aatccttgag agttttcgcc ctgaagagcg ttttccaatg atgagcactt
     1201 tcaaagttct gctatgtgga gcagtattat cccgtgtaga tgcggggcaa gagcaactcg
     1261 gacgacgaat acactattcg cagaatgact tggttgaata ctccccagtg acagaaaagc
     1321 accttacgga cggaatgacg gtaagagaat tatgtagtgc cgccataacg atgagtgata
     1381 acactgcggc gaacttactt ctgacaacca tcggtggacc gaaggaatta accgcttttt
     1441 tgcacaatat gggagaccat gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag
     1501 ccataccaaa cgacgagcga gacaccacaa tgcctgcggc aatggcaaca acattacgca
     1561 aactattaac tggcgaacta cttactctgg cttcacggca acaattaata gactggcttg
     1621 aagcggataa agttgcagga ccactactgc gttcggcact tcctgctggc tggtttattg
     1681 ctgataaatc tggggcagga gagcgtggtt cacggggtat cattgccgca cttggaccag
     1741 atggtaagcc ttcccgtatc gtagttatct acacgacggg tagtcaggca actatggacg
     1801 aacgaaatag acagattgct gaaatagggg cttcactgat taagcattgg taaaccgata
     1861 caattaaagg ctccttttgg agcctttttt tttggacgga ccggtagaaa agatcaaagg
     1921 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1981 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2041 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2101 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2161 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2221 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2281 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2341 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2401 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2461 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2521 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2581 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2641 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2701 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     2761 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     2821 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2881 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2941 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3001 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3061 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3121 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg taatgatacc
     3181 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     3241 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     3301 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gtt
//
