LOCUS       sequence-177403-        3307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term 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 3307)
  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 3307)
  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-FLAGNDHFR
FEATURES             Location/Qualifiers
     source          1..3307
                     /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"
     CDS             164..184
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(739..757)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      753..800
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        865..956
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1821..1860
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1891..2479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2380..2399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2633..2650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2665..2804
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2730..2749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2890..2912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2906..3097)
                     /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 agggatctgg tggtggtggt ggtgaaaacc tgtacttcca
      181 gggatccatc agtctgattg cggcgctagc ggtagatcgc gttatcggca tggaaaacgc
      241 catgccatgg aacctgcctg ccgatctcgc ctggtttaaa cgcaacacct taaataaacc
      301 cgtgattatg gggcgccata cctgggaatc aatcggtagg cctttgcccg gccgcaaaaa
      361 tattatcctc agcagtcaac ccgggaccga tgatcgggtt acctgggtta aatcggtcga
      421 cgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg gtgattggcg gcggacgcgt
      481 ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg acgcatatcg atgcagaagt
      541 ggaaggcgac acccattttc cggattacga gccggatgac tgggaatcgg tattcagcga
      601 attccacgat gctgatgcgc agaactcgca tagctattgt ttcgaaatcc tcgagcgtcg
      661 tgccggctaa tgtacaacgc gtgcgcgctg atccggctgc taacaaagcc cgaaaggaag
      721 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      781 gggtcttgag gggttttttg ctgaaaggag gaactatatc gggattagcc cgcctaatga
      841 gcgggctttt ttttaattcc cctatttgtt tatttttcta aatacattca aatatgtatc
      901 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1021 ttgctcaccc agaaacgctc gtgaaagtaa aagacgcaga ggaccaattg ggggcacgag
     1081 tgggatacat agaactggac ttgaatagcg gtaaaatcct tgagagtttt cgccctgaag
     1141 agcgttttcc aatgatgagc actttcaaag ttctgctatg tggagcagta ttatcccgtg
     1201 tagatgcggg gcaagagcaa ctcggacgac gaatacacta ttcgcagaat gacttggttg
     1261 aatactcccc agtgacagaa aagcacctta cggacggaat gacggtaaga gaattatgta
     1321 gtgccgccat aacgatgagt gataacactg cggcgaactt acttctgaca accatcggtg
     1381 gaccgaagga attaaccgct tttttgcaca atatgggaga ccatgtaact cgccttgacc
     1441 gttgggaacc agaactgaat gaagccatac caaacgacga gcgagacacc acaatgcctg
     1501 cggcaatggc aacaacatta cgcaaactat taactggcga actacttact ctggcttcac
     1561 ggcaacaatt aatagactgg cttgaagcgg ataaagttgc aggaccacta ctgcgttcgg
     1621 cacttcctgc tggctggttt attgctgata aatctggggc aggagagcgt ggttcacggg
     1681 gtatcattgc cgcacttgga ccagatggta agccttcccg tatcgtagtt atctacacga
     1741 cgggtagtca ggcaactatg gacgaacgaa atagacagat tgctgaaata ggggcttcac
     1801 tgattaagca ttggtaaacc gatacaatta aaggctcctt ttggagcctt tttttttgga
     1861 cggaccggta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1921 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1981 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2041 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2101 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2161 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2221 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2281 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2341 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2401 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2461 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2521 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2581 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2641 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2701 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2761 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2821 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2881 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2941 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3001 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3061 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3121 ttctgttcat ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3181 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3241 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3301 aggtgtt
//
