LOCUS       sequence-177464-        3406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 1 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 3406)
  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 3406)
  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: pST50Trc1-HPCHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3406
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..184
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             194..211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             239..259
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(838..856)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      852..899
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        964..1055
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1920..1959
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1990..2578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2479..2498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2732..2749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2764..2903
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2829..2848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2989..3011
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3005..3196)
                     /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 attcgctagc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatgtctga agatcaggta gatccacgtt taatcgacgg
      181 taaaggatct agccatcatc atcatcatca cagcagcgga tctggtggtg gtggtggtga
      241 aaacctgtac ttccagggat ccatcagtct gattgcggcg ctagcggtag atcgcgttat
      301 cggcatggaa aacgccatgc catggaacct gcctgccgat ctcgcctggt ttaaacgcaa
      361 caccttaaat aaacccgtga ttatggggcg ccatacctgg gaatcaatcg gtaggccttt
      421 gcccggccgc aaaaatatta tcctcagcag tcaacccggg accgatgatc gggttacctg
      481 ggttaaatcg gtcgacgaag ccatcgcggc gtgtggtgac gtaccagaaa tcatggtgat
      541 tggcggcgga cgcgtttatg aacagttctt gccaaaagcg caaaaactgt atctgacgca
      601 tatcgatgca gaagtggaag gcgacaccca ttttccggat tacgagccgg atgactggga
      661 atcggtattc agcgaattcc acgatgctga tgcgcagaac tcgcatagct attgtttcga
      721 aatcctcgag cgtcgtgccg gctaatgtac aagatccaga tctaagcttg gtaccacgcg
      781 tgcgcgctga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      841 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      901 tgaaaggagg aactatatcg ggattagccc gcctaatgag cgggcttttt tttaattccc
      961 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1021 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1081 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctcg
     1141 tgaaagtaaa agacgcagag gaccaattgg gggcacgagt gggatacata gaactggact
     1201 tgaatagcgg taaaatcctt gagagttttc gccctgaaga gcgttttcca atgatgagca
     1261 ctttcaaagt tctgctatgt ggagcagtat tatcccgtgt agatgcgggg caagagcaac
     1321 tcggacgacg aatacactat tcgcagaatg acttggttga atactcccca gtgacagaaa
     1381 agcaccttac ggacggaatg acggtaagag aattatgtag tgccgccata acgatgagtg
     1441 ataacactgc ggcgaactta cttctgacaa ccatcggtgg accgaaggaa ttaaccgctt
     1501 ttttgcacaa tatgggagac catgtaactc gccttgaccg ttgggaacca gaactgaatg
     1561 aagccatacc aaacgacgag cgagacacca caatgcctgc ggcaatggca acaacattac
     1621 gcaaactatt aactggcgaa ctacttactc tggcttcacg gcaacaatta atagactggc
     1681 ttgaagcgga taaagttgca ggaccactac tgcgttcggc acttcctgct ggctggttta
     1741 ttgctgataa atctggggca ggagagcgtg gttcacgggg tatcattgcc gcacttggac
     1801 cagatggtaa gccttcccgt atcgtagtta tctacacgac gggtagtcag gcaactatgg
     1861 acgaacgaaa tagacagatt gctgaaatag gggcttcact gattaagcat tggtaaaccg
     1921 atacaattaa aggctccttt tggagccttt ttttttggac ggaccggtag aaaagatcaa
     1981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2101 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2641 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2701 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2761 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     2821 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     2881 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     2941 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3001 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3061 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3121 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3181 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg gggtaatgat
     3241 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3301 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3361 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgtt
//
