LOCUS       sequence-177412-        3265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 transfer plasmid for pST44 polycistronic plasmid suite; 
            C-term 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 3265)
  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 3265)
  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-DHFRFLAG
FEATURES             Location/Qualifiers
     source          1..3265
                     /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             602..625
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(697..715)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      711..758
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        823..914
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1779..1818
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1849..2437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2338..2357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2591..2608
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2623..2762
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2688..2707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2848..2870
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2864..3055)
                     /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 atatgggatc
      121 catcagtctg attgcggcgc tagcggtaga tcgcgttatc ggcatggaaa acgccatgcc
      181 atggaacctg cctgccgatc tcgcctggtt taaacgcaac accttaaata aacccgtgat
      241 tatggggcgc catacctggg aatcaatcgg taggcctttg cccggccgca aaaatattat
      301 cctcagcagt caacccggga ccgatgatcg ggttacctgg gttaaatcgg tcgacgaagc
      361 catcgcggcg tgtggtgacg taccagaaat catggtgatt ggcggcggac gcgtttatga
      421 acagttcttg ccaaaagcgc aaaaactgta tctgacgcat atcgatgcag aagtggaagg
      481 cgacacccat tttccggatt acgagccgga tgactgggaa tcggtattca gcgaattcca
      541 cgatgctgat gcgcagaact cgcatagcta ttgtttcgaa atcctcgagc gtcgtgccgg
      601 cgactacaag gacgatgatg acaagtaatg tacaacgcgt gcgcgctgat ccggctgcta
      661 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
      721 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatcgg
      781 gattagcccg cctaatgagc gggctttttt ttaattcccc tatttgttta tttttctaaa
      841 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      901 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      961 cattttgcct tcctgttttt gctcacccag aaacgctcgt gaaagtaaaa gacgcagagg
     1021 accaattggg ggcacgagtg ggatacatag aactggactt gaatagcggt aaaatccttg
     1081 agagttttcg ccctgaagag cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg
     1141 gagcagtatt atcccgtgta gatgcggggc aagagcaact cggacgacga atacactatt
     1201 cgcagaatga cttggttgaa tactccccag tgacagaaaa gcaccttacg gacggaatga
     1261 cggtaagaga attatgtagt gccgccataa cgatgagtga taacactgcg gcgaacttac
     1321 ttctgacaac catcggtgga ccgaaggaat taaccgcttt tttgcacaat atgggagacc
     1381 atgtaactcg ccttgaccgt tgggaaccag aactgaatga agccatacca aacgacgagc
     1441 gagacaccac aatgcctgcg gcaatggcaa caacattacg caaactatta actggcgaac
     1501 tacttactct ggcttcacgg caacaattaa tagactggct tgaagcggat aaagttgcag
     1561 gaccactact gcgttcggca cttcctgctg gctggtttat tgctgataaa tctggggcag
     1621 gagagcgtgg ttcacggggt atcattgccg cacttggacc agatggtaag ccttcccgta
     1681 tcgtagttat ctacacgacg ggtagtcagg caactatgga cgaacgaaat agacagattg
     1741 ctgaaatagg ggcttcactg attaagcatt ggtaaaccga tacaattaaa ggctcctttt
     1801 ggagcctttt tttttggacg gaccggtaga aaagatcaaa ggatcttctt gagatccttt
     1861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1981 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2521 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2581 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2641 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2701 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2761 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2821 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2881 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     2941 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3001 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3061 cctccgtgta agggggattt ctgttcatgg ggtaatgata ccgatgaaac gagagaggat
     3121 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3181 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     3241 cttcgttaat acagatgtag gtgtt
//
