LOCUS       sequence-177385-        3319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 3 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 3319)
  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 3319)
  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-FLAGNDHFR
FEATURES             Location/Qualifiers
     source          1..3319
                     /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             125..148
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             170..190
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(751..769)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      765..812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        877..968
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1833..1872
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1903..2491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2392..2411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2645..2662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2677..2816
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2742..2761)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2902..2924
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2918..3109)
                     /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 gggagactac aaggacgatg atgacaaggg atctggtggt ggtggtggtg aaaacctgta
      181 cttccaggga tccatcagtc tgattgcggc gctagcggta gatcgcgtta tcggcatgga
      241 aaacgccatg ccatggaacc tgcctgccga tctcgcctgg tttaaacgca acaccttaaa
      301 taaacccgtg attatggggc gccatacctg ggaatcaatc ggtaggcctt tgcccggccg
      361 caaaaatatt atcctcagca gtcaacccgg gaccgatgat cgggttacct gggttaaatc
      421 ggtcgacgaa gccatcgcgg cgtgtggtga cgtaccagaa atcatggtga ttggcggcgg
      481 acgcgtttat gaacagttct tgccaaaagc gcaaaaactg tatctgacgc atatcgatgc
      541 agaagtggaa ggcgacaccc attttccgga ttacgagccg gatgactggg aatcggtatt
      601 cagcgaattc cacgatgctg atgcgcagaa ctcgcatagc tattgtttcg aaatcctcga
      661 gcgtcgtgcc ggctaatgta caggtaccac gcgtgcgcgc tgatccggct gctaacaaag
      721 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      781 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tcgggattag
      841 cccgcctaat gagcgggctt ttttttaatt cccctatttg tttatttttc taaatacatt
      901 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      961 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1021 gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca gaggaccaat
     1081 tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc cttgagagtt
     1141 ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta tgtggagcag
     1201 tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac tattcgcaga
     1261 atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga atgacggtaa
     1321 gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac ttacttctga
     1381 caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga gaccatgtaa
     1441 ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac gagcgagaca
     1501 ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc gaactactta
     1561 ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt gcaggaccac
     1621 tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg gcaggagagc
     1681 gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc cgtatcgtag
     1741 ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag attgctgaaa
     1801 taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc ttttggagcc
     1861 tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2041 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2281 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2581 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2641 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2701 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     2761 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2821 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2881 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2941 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3001 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3061 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3121 tgtaaggggg atttctgttc atggggtaat gataccgatg aaacgagaga ggatgctcac
     3181 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     3241 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3301 taatacagat gtaggtgtt
//
