LOCUS       sequence-177393-        3277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 3 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 3277)
  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 3277)
  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: pST50Trc3-DHFRFLAG
FEATURES             Location/Qualifiers
     source          1..3277
                     /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             89..111
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             608..631
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(709..727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      723..770
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        835..926
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1791..1830
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1861..2449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2350..2369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2603..2620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2635..2774
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2700..2719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2860..2882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2876..3067)
                     /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 tcaataattt tgtttaactt taagaaggag atatacatat
      121 gggatccatc agtctgattg cggcgctagc ggtagatcgc gttatcggca tggaaaacgc
      181 catgccatgg aacctgcctg ccgatctcgc ctggtttaaa cgcaacacct taaataaacc
      241 cgtgattatg gggcgccata cctgggaatc aatcggtagg cctttgcccg gccgcaaaaa
      301 tattatcctc agcagtcaac ccgggaccga tgatcgggtt acctgggtta aatcggtcga
      361 cgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg gtgattggcg gcggacgcgt
      421 ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg acgcatatcg atgcagaagt
      481 ggaaggcgac acccattttc cggattacga gccggatgac tgggaatcgg tattcagcga
      541 attccacgat gctgatgcgc agaactcgca tagctattgt ttcgaaatcc tcgagcgtcg
      601 tgccggcgac tacaaggacg atgatgacaa gtaatgtaca ggtaccacgc gtgcgcgctg
      661 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      721 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      781 gaactatatc gggattagcc cgcctaatga gcgggctttt ttttaattcc cctatttgtt
      841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      901 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      961 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctc gtgaaagtaa
     1021 aagacgcaga ggaccaattg ggggcacgag tgggatacat agaactggac ttgaatagcg
     1081 gtaaaatcct tgagagtttt cgccctgaag agcgttttcc aatgatgagc actttcaaag
     1141 ttctgctatg tggagcagta ttatcccgtg tagatgcggg gcaagagcaa ctcggacgac
     1201 gaatacacta ttcgcagaat gacttggttg aatactcccc agtgacagaa aagcacctta
     1261 cggacggaat gacggtaaga gaattatgta gtgccgccat aacgatgagt gataacactg
     1321 cggcgaactt acttctgaca accatcggtg gaccgaagga attaaccgct tttttgcaca
     1381 atatgggaga ccatgtaact cgccttgacc gttgggaacc agaactgaat gaagccatac
     1441 caaacgacga gcgagacacc acaatgcctg cggcaatggc aacaacatta cgcaaactat
     1501 taactggcga actacttact ctggcttcac ggcaacaatt aatagactgg cttgaagcgg
     1561 ataaagttgc aggaccacta ctgcgttcgg cacttcctgc tggctggttt attgctgata
     1621 aatctggggc aggagagcgt ggttcacggg gtatcattgc cgcacttgga ccagatggta
     1681 agccttcccg tatcgtagtt atctacacga cgggtagtca ggcaactatg gacgaacgaa
     1741 atagacagat tgctgaaata ggggcttcac tgattaagca ttggtaaacc gatacaatta
     1801 aaggctcctt ttggagcctt tttttttgga cggaccggta gaaaagatca aaggatcttc
     1861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1981 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2521 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2581 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     2641 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     2701 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2761 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2821 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2881 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     2941 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3001 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3061 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     3121 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3181 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3241 tcaatgccag cgcttcgtta atacagatgt aggtgtt
//
