LOCUS       sequence-177409-        3259 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 3259)
  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 3259)
  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-DHFRHIS
FEATURES             Location/Qualifiers
     source          1..3259
                     /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..619
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(691..709)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      705..752
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        817..908
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1773..1812
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1843..2431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2332..2351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2585..2602
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2617..2756
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2682..2701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2842..2864
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2858..3049)
                     /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 ccaccaccac caccaccact aatgtacaac gcgtgcgcgc tgatccggct gctaacaaag
      661 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      721 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tcgggattag
      781 cccgcctaat gagcgggctt ttttttaatt cccctatttg tttatttttc taaatacatt
      841 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      901 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      961 gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca gaggaccaat
     1021 tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc cttgagagtt
     1081 ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta tgtggagcag
     1141 tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac tattcgcaga
     1201 atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga atgacggtaa
     1261 gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac ttacttctga
     1321 caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga gaccatgtaa
     1381 ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac gagcgagaca
     1441 ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc gaactactta
     1501 ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt gcaggaccac
     1561 tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg gcaggagagc
     1621 gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc cgtatcgtag
     1681 ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag attgctgaaa
     1741 taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc ttttggagcc
     1801 tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1981 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2461 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2521 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2581 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2641 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     2701 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2761 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2821 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2881 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2941 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3001 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3061 tgtaaggggg atttctgttc atggggtaat gataccgatg aaacgagaga ggatgctcac
     3121 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     3181 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3241 taatacagat gtaggtgtt
//
