LOCUS       sequence-177500-        3382 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 3382)
  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 3382)
  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-HPCHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3382
                     /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             137..172
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             182..199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             227..247
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(814..832)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      828..875
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        940..1031
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1896..1935
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1966..2554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2455..2474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2708..2725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2740..2879
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2805..2824)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2965..2987
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2981..3172)
                     /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 atgtctgaag atcaggtaga tccacgttta atcgacggta aaggatctag
      181 ccatcatcat catcatcaca gcagcggatc tggtggtggt ggtggtgaaa acctgtactt
      241 ccagggatcc atcagtctga ttgcggcgct agcggtagat cgcgttatcg gcatggaaaa
      301 cgccatgcca tggaacctgc ctgccgatct cgcctggttt aaacgcaaca ccttaaataa
      361 acccgtgatt atggggcgcc atacctggga atcaatcggt aggcctttgc ccggccgcaa
      421 aaatattatc ctcagcagtc aacccgggac cgatgatcgg gttacctggg ttaaatcggt
      481 cgacgaagcc atcgcggcgt gtggtgacgt accagaaatc atggtgattg gcggcggacg
      541 cgtttatgaa cagttcttgc caaaagcgca aaaactgtat ctgacgcata tcgatgcaga
      601 agtggaaggc gacacccatt ttccggatta cgagccggat gactgggaat cggtattcag
      661 cgaattccac gatgctgatg cgcagaactc gcatagctat tgtttcgaaa tcctcgagcg
      721 tcgtgccggc taatgtacaa agcttggtac cacgcgtgcg cgctgatccg gctgctaaca
      781 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
      841 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatcgggat
      901 tagcccgcct aatgagcggg ctttttttta attcccctat ttgtttattt ttctaaatac
      961 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1021 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1081 tttgccttcc tgtttttgct cacccagaaa cgctcgtgaa agtaaaagac gcagaggacc
     1141 aattgggggc acgagtggga tacatagaac tggacttgaa tagcggtaaa atccttgaga
     1201 gttttcgccc tgaagagcgt tttccaatga tgagcacttt caaagttctg ctatgtggag
     1261 cagtattatc ccgtgtagat gcggggcaag agcaactcgg acgacgaata cactattcgc
     1321 agaatgactt ggttgaatac tccccagtga cagaaaagca ccttacggac ggaatgacgg
     1381 taagagaatt atgtagtgcc gccataacga tgagtgataa cactgcggcg aacttacttc
     1441 tgacaaccat cggtggaccg aaggaattaa ccgctttttt gcacaatatg ggagaccatg
     1501 taactcgcct tgaccgttgg gaaccagaac tgaatgaagc cataccaaac gacgagcgag
     1561 acaccacaat gcctgcggca atggcaacaa cattacgcaa actattaact ggcgaactac
     1621 ttactctggc ttcacggcaa caattaatag actggcttga agcggataaa gttgcaggac
     1681 cactactgcg ttcggcactt cctgctggct ggtttattgc tgataaatct ggggcaggag
     1741 agcgtggttc acggggtatc attgccgcac ttggaccaga tggtaagcct tcccgtatcg
     1801 tagttatcta cacgacgggt agtcaggcaa ctatggacga acgaaataga cagattgctg
     1861 aaataggggc ttcactgatt aagcattggt aaaccgatac aattaaaggc tccttttgga
     1921 gccttttttt ttggacggac cggtagaaaa gatcaaagga tcttcttgag atcctttttt
     1981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2101 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2701 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2761 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     2821 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     2881 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2941 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3001 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     3061 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     3121 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     3181 ccgtgtaagg gggatttctg ttcatggggt aatgataccg atgaaacgag agaggatgct
     3241 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     3301 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     3361 cgttaataca gatgtaggtg tt
//
