LOCUS       sequence-177498-        3370 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 3370)
  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 3370)
  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-FLAGHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3370
                     /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             170..187
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             215..235
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(802..820)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      816..863
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        928..1019
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1884..1923
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1954..2542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2443..2462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2696..2713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2728..2867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2793..2812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2953..2975
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2969..3160)
                     /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 ggatctagcc atcatcatca
      181 tcatcacagc agcggatctg gtggtggtgg tggtgaaaac ctgtacttcc agggatccat
      241 cagtctgatt gcggcgctag cggtagatcg cgttatcggc atggaaaacg ccatgccatg
      301 gaacctgcct gccgatctcg cctggtttaa acgcaacacc ttaaataaac ccgtgattat
      361 ggggcgccat acctgggaat caatcggtag gcctttgccc ggccgcaaaa atattatcct
      421 cagcagtcaa cccgggaccg atgatcgggt tacctgggtt aaatcggtcg acgaagccat
      481 cgcggcgtgt ggtgacgtac cagaaatcat ggtgattggc ggcggacgcg tttatgaaca
      541 gttcttgcca aaagcgcaaa aactgtatct gacgcatatc gatgcagaag tggaaggcga
      601 cacccatttt ccggattacg agccggatga ctgggaatcg gtattcagcg aattccacga
      661 tgctgatgcg cagaactcgc atagctattg tttcgaaatc ctcgagcgtc gtgccggcta
      721 atgtacaaag cttggtacca cgcgtgcgcg ctgatccggc tgctaacaaa gcccgaaagg
      781 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      841 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atcgggatta gcccgcctaa
      901 tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt
      961 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1021 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1081 tttttgctca cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa ttgggggcac
     1141 gagtgggata catagaactg gacttgaata gcggtaaaat ccttgagagt tttcgccctg
     1201 aagagcgttt tccaatgatg agcactttca aagttctgct atgtggagca gtattatccc
     1261 gtgtagatgc ggggcaagag caactcggac gacgaataca ctattcgcag aatgacttgg
     1321 ttgaatactc cccagtgaca gaaaagcacc ttacggacgg aatgacggta agagaattat
     1381 gtagtgccgc cataacgatg agtgataaca ctgcggcgaa cttacttctg acaaccatcg
     1441 gtggaccgaa ggaattaacc gcttttttgc acaatatggg agaccatgta actcgccttg
     1501 accgttggga accagaactg aatgaagcca taccaaacga cgagcgagac accacaatgc
     1561 ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg cgaactactt actctggctt
     1621 cacggcaaca attaatagac tggcttgaag cggataaagt tgcaggacca ctactgcgtt
     1681 cggcacttcc tgctggctgg tttattgctg ataaatctgg ggcaggagag cgtggttcac
     1741 ggggtatcat tgccgcactt ggaccagatg gtaagccttc ccgtatcgta gttatctaca
     1801 cgacgggtag tcaggcaact atggacgaac gaaatagaca gattgctgaa ataggggctt
     1861 cactgattaa gcattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt
     1921 ggacggaccg gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2101 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2701 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     2761 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2821 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     2881 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2941 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3001 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3061 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3121 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     3181 gatttctgtt catggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3241 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3301 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3361 tgtaggtgtt
//
