LOCUS       sequence-177362-        3298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term non-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 3298)
  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 3298)
  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-FLAGDHFR
FEATURES             Location/Qualifiers
     source          1..3298
                     /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"
     primer_bind     complement(730..748)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      744..791
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        856..947
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1812..1851
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1882..2470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2371..2390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2624..2641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2656..2795
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2721..2740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2881..2903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2897..3088)
                     /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 ggatccatca gtctgattgc
      181 ggcgctagcg gtagatcgcg ttatcggcat ggaaaacgcc atgccatgga acctgcctgc
      241 cgatctcgcc tggtttaaac gcaacacctt aaataaaccc gtgattatgg ggcgccatac
      301 ctgggaatca atcggtaggc ctttgcccgg ccgcaaaaat attatcctca gcagtcaacc
      361 cgggaccgat gatcgggtta cctgggttaa atcggtcgac gaagccatcg cggcgtgtgg
      421 tgacgtacca gaaatcatgg tgattggcgg cggacgcgtt tatgaacagt tcttgccaaa
      481 agcgcaaaaa ctgtatctga cgcatatcga tgcagaagtg gaaggcgaca cccattttcc
      541 ggattacgag ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca
      601 gaactcgcat agctattgtt tcgaaatcct cgagcgtcgt gccggctaat gtacaaagct
      661 tggtaccacg cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
      721 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
      781 ggggtttttt gctgaaagga ggaactatat cgggattagc ccgcctaatg agcgggcttt
      841 tttttaattc ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1021 cagaaacgct cgtgaaagta aaagacgcag aggaccaatt gggggcacga gtgggataca
     1081 tagaactgga cttgaatagc ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc
     1141 caatgatgag cactttcaaa gttctgctat gtggagcagt attatcccgt gtagatgcgg
     1201 ggcaagagca actcggacga cgaatacact attcgcagaa tgacttggtt gaatactccc
     1261 cagtgacaga aaagcacctt acggacggaa tgacggtaag agaattatgt agtgccgcca
     1321 taacgatgag tgataacact gcggcgaact tacttctgac aaccatcggt ggaccgaagg
     1381 aattaaccgc ttttttgcac aatatgggag accatgtaac tcgccttgac cgttgggaac
     1441 cagaactgaa tgaagccata ccaaacgacg agcgagacac cacaatgcct gcggcaatgg
     1501 caacaacatt acgcaaacta ttaactggcg aactacttac tctggcttca cggcaacaat
     1561 taatagactg gcttgaagcg gataaagttg caggaccact actgcgttcg gcacttcctg
     1621 ctggctggtt tattgctgat aaatctgggg caggagagcg tggttcacgg ggtatcattg
     1681 ccgcacttgg accagatggt aagccttccc gtatcgtagt tatctacacg acgggtagtc
     1741 aggcaactat ggacgaacga aatagacaga ttgctgaaat aggggcttca ctgattaagc
     1801 attggtaaac cgatacaatt aaaggctcct tttggagcct ttttttttgg acggaccggt
     1861 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1921 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1981 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2041 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2101 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2161 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2221 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2281 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2341 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2401 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2461 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2521 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2581 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2641 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2701 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     2761 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     2821 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2881 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     2941 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3001 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3061 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3121 tggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3181 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3241 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgtt
//
