LOCUS       sequence-177367-        3337 bp    DNA     circular SYN 15-DEC-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 3337)
  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 3337)
  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-FLAGNDHFR
FEATURES             Location/Qualifiers
     source          1..3337
                     /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..160
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             182..202
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(769..787)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      783..830
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        895..986
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1851..1890
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1921..2509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2410..2429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2663..2680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2695..2834
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2760..2779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2920..2942
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2936..3127)
                     /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 atgggagact acaaggacga tgatgacaag ggatctggtg gtggtggtgg
      181 tgaaaacctg tacttccagg gatccatcag tctgattgcg gcgctagcgg tagatcgcgt
      241 tatcggcatg gaaaacgcca tgccatggaa cctgcctgcc gatctcgcct ggtttaaacg
      301 caacacctta aataaacccg tgattatggg gcgccatacc tgggaatcaa tcggtaggcc
      361 tttgcccggc cgcaaaaata ttatcctcag cagtcaaccc gggaccgatg atcgggttac
      421 ctgggttaaa tcggtcgacg aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt
      481 gattggcggc ggacgcgttt atgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac
      541 gcatatcgat gcagaagtgg aaggcgacac ccattttccg gattacgagc cggatgactg
      601 ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag aactcgcata gctattgttt
      661 cgaaatcctc gagcgtcgtg ccggctaatg tacaaagctt ggtaccacgc gtgcgcgctg
      721 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      781 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      841 gaactatatc gggattagcc cgcctaatga gcgggctttt ttttaattcc cctatttgtt
      901 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      961 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1021 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctc gtgaaagtaa
     1081 aagacgcaga ggaccaattg ggggcacgag tgggatacat agaactggac ttgaatagcg
     1141 gtaaaatcct tgagagtttt cgccctgaag agcgttttcc aatgatgagc actttcaaag
     1201 ttctgctatg tggagcagta ttatcccgtg tagatgcggg gcaagagcaa ctcggacgac
     1261 gaatacacta ttcgcagaat gacttggttg aatactcccc agtgacagaa aagcacctta
     1321 cggacggaat gacggtaaga gaattatgta gtgccgccat aacgatgagt gataacactg
     1381 cggcgaactt acttctgaca accatcggtg gaccgaagga attaaccgct tttttgcaca
     1441 atatgggaga ccatgtaact cgccttgacc gttgggaacc agaactgaat gaagccatac
     1501 caaacgacga gcgagacacc acaatgcctg cggcaatggc aacaacatta cgcaaactat
     1561 taactggcga actacttact ctggcttcac ggcaacaatt aatagactgg cttgaagcgg
     1621 ataaagttgc aggaccacta ctgcgttcgg cacttcctgc tggctggttt attgctgata
     1681 aatctggggc aggagagcgt ggttcacggg gtatcattgc cgcacttgga ccagatggta
     1741 agccttcccg tatcgtagtt atctacacga cgggtagtca ggcaactatg gacgaacgaa
     1801 atagacagat tgctgaaata ggggcttcac tgattaagca ttggtaaacc gatacaatta
     1861 aaggctcctt ttggagcctt tttttttgga cggaccggta gaaaagatca aaggatcttc
     1921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2041 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2281 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2521 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2581 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2641 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     2701 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     2761 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2821 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2881 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2941 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3001 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3061 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3121 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     3181 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3241 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3301 tcaatgccag cgcttcgtta atacagatgt aggtgtt
//
