LOCUS       sequence-177398-        3268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3268)
  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 3268)
  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: pST50Trc4-FLAGDHFR
FEATURES             Location/Qualifiers
     source          1..3268
                     /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             83..105
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             119..142
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(700..718)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      714..761
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        826..917
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1782..1821
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1852..2440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2341..2360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2594..2611
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2626..2765
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2691..2710)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2851..2873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2867..3058)
                     /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 tccggaaata attttgttta actttaagaa ggagatatac atatgggaga
      121 ctacaaggac gatgatgaca agggatccat cagtctgatt gcggcgctag cggtagatcg
      181 cgttatcggc atggaaaacg ccatgccatg gaacctgcct gccgatctcg cctggtttaa
      241 acgcaacacc ttaaataaac ccgtgattat ggggcgccat acctgggaat caatcggtag
      301 gcctttgccc ggccgcaaaa atattatcct cagcagtcaa cccgggaccg atgatcgggt
      361 tacctgggtt aaatcggtcg acgaagccat cgcggcgtgt ggtgacgtac cagaaatcat
      421 ggtgattggc ggcggacgcg tttatgaaca gttcttgcca aaagcgcaaa aactgtatct
      481 gacgcatatc gatgcagaag tggaaggcga cacccatttt ccggattacg agccggatga
      541 ctgggaatcg gtattcagcg aattccacga tgctgatgcg cagaactcgc atagctattg
      601 tttcgaaatc ctcgagcgtc gtgccggcta atgtacaacg cgtgcgcgct gatccggctg
      661 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
      721 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
      781 cgggattagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt ttatttttct
      841 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      901 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      961 cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta aaagacgcag
     1021 aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc ggtaaaatcc
     1081 ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa gttctgctat
     1141 gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga cgaatacact
     1201 attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt acggacggaa
     1261 tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact gcggcgaact
     1321 tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac aatatgggag
     1381 accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata ccaaacgacg
     1441 agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta ttaactggcg
     1501 aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
     1561 caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
     1621 caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
     1681 gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
     1741 ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
     1801 tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt cttgagatcc
     1861 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1921 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1981 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2041 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2101 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2161 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2221 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2281 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2341 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2401 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2461 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2521 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2581 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2641 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     2701 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2761 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2821 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2881 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     2941 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3001 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3061 atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag
     3121 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     3181 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     3241 gcgcttcgtt aatacagatg taggtgtt
//
