LOCUS       sequence-177472-        3300 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 3300)
  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 3300)
  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-HISDHFR
FEATURES             Location/Qualifiers
     source          1..3300
                     /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             143..160
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(736..754)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      750..797
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        858..949
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1814..1853
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1884..2472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2373..2392
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2626..2643
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2658..2797
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2723..2742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2883..2905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2899..3090)
                     /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 atgggcagca gccatcatca tcatcatcac agcagcggat ccatcagtct
      181 gattgcggcg ctagcggtag atcgcgttat cggcatggaa aacgccatgc catggaacct
      241 gcctgccgat ctcgcctggt ttaaacgcaa caccttaaat aaacccgtga ttatggggcg
      301 ccatacctgg gaatcaatcg gtaggccttt gcccggccgc aaaaatatta tcctcagcag
      361 tcaacccggg accgatgatc gggttacctg ggttaaatcg gtcgacgaag ccatcgcggc
      421 gtgtggtgac gtaccagaaa tcatggtgat tggcggcgga cgcgtttatg aacagttctt
      481 gccaaaagcg caaaaactgt atctgacgca tatcgatgca gaagtggaag gcgacaccca
      541 ttttccggat tacgagccgg atgactggga atcggtattc agcgaattcc acgatgctga
      601 tgcgcagaac tcgcatagct attgtttcga aatcctcgag cgtcgtgccg gctaatgtac
      661 aaagcttggt accacgcgtg cgcgctgatc cggctgctaa caaagcccga aaggaagctg
      721 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
      781 tcttgagggg ttttttgctg aaaggaggaa ctatatatta gcccgcctaa tgagcgggct
      841 tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      901 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      961 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1021 cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa ttgggggcac gagtgggata
     1081 catagaactg gacttgaata gcggtaaaat ccttgagagt tttcgccctg aagagcgttt
     1141 tccaatgatg agcactttca aagttctgct atgtggagca gtattatccc gtgtagatgc
     1201 ggggcaagag caactcggac gacgaataca ctattcgcag aatgacttgg ttgaatactc
     1261 cccagtgaca gaaaagcacc ttacggacgg aatgacggta agagaattat gtagtgccgc
     1321 cataacgatg agtgataaca ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa
     1381 ggaattaacc gcttttttgc acaatatggg agaccatgta actcgccttg accgttggga
     1441 accagaactg aatgaagcca taccaaacga cgagcgagac accacaatgc ctgcggcaat
     1501 ggcaacaaca ttacgcaaac tattaactgg cgaactactt actctggctt cacggcaaca
     1561 attaatagac tggcttgaag cggataaagt tgcaggacca ctactgcgtt cggcacttcc
     1621 tgctggctgg tttattgctg ataaatctgg ggcaggagag cgtggttcac ggggtatcat
     1681 tgccgcactt ggaccagatg gtaagccttc ccgtatcgta gttatctaca cgacgggtag
     1741 tcaggcaact atggacgaac gaaatagaca gattgctgaa ataggggctt cactgattaa
     1801 gcattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt ggacggaccg
     1861 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1921 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1981 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2041 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2101 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2161 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2221 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2281 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2341 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2401 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2461 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2521 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2581 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2641 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2701 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     2761 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     2821 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2881 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     2941 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3001 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3061 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     3121 catggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     3181 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     3241 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
//
