LOCUS       sequence-177388-        3355 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Position 3 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term cleavable double 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 3355)
  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 3355)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-STRHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3355
                     /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             128..151
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             161..178
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             206..226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(787..805)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      801..848
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        913..1004
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1869..1908
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1939..2527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2428..2447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2681..2698
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2713..2852
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2778..2797)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2938..2960
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2954..3145)
                     /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 ggcaagctgg agccacccgc agttcgagaa aggatctagc catcatcatc atcatcacag
      181 cagcggatct ggtggtggtg gtggtgaaaa cctgtacttc cagggatcca tcagtctgat
      241 tgcggcgcta gcggtagatc gcgttatcgg catggaaaac gccatgccat ggaacctgcc
      301 tgccgatctc gcctggttta aacgcaacac cttaaataaa cccgtgatta tggggcgcca
      361 tacctgggaa tcaatcggta ggcctttgcc cggccgcaaa aatattatcc tcagcagtca
      421 acccgggacc gatgatcggg ttacctgggt taaatcggtc gacgaagcca tcgcggcgtg
      481 tggtgacgta ccagaaatca tggtgattgg cggcggacgc gtttatgaac agttcttgcc
      541 aaaagcgcaa aaactgtatc tgacgcatat cgatgcagaa gtggaaggcg acacccattt
      601 tccggattac gagccggatg actgggaatc ggtattcagc gaattccacg atgctgatgc
      661 gcagaactcg catagctatt gtttcgaaat cctcgagcgt cgtgccggct aatgtacagg
      721 taccacgcgt gcgcgctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      841 gttttttgct gaaaggagga actatatcgg gattagcccg cctaatgagc gggctttttt
      901 ttaattcccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      961 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1021 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1081 aaacgctcgt gaaagtaaaa gacgcagagg accaattggg ggcacgagtg ggatacatag
     1141 aactggactt gaatagcggt aaaatccttg agagttttcg ccctgaagag cgttttccaa
     1201 tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt atcccgtgta gatgcggggc
     1261 aagagcaact cggacgacga atacactatt cgcagaatga cttggttgaa tactccccag
     1321 tgacagaaaa gcaccttacg gacggaatga cggtaagaga attatgtagt gccgccataa
     1381 cgatgagtga taacactgcg gcgaacttac ttctgacaac catcggtgga ccgaaggaat
     1441 taaccgcttt tttgcacaat atgggagacc atgtaactcg ccttgaccgt tgggaaccag
     1501 aactgaatga agccatacca aacgacgagc gagacaccac aatgcctgcg gcaatggcaa
     1561 caacattacg caaactatta actggcgaac tacttactct ggcttcacgg caacaattaa
     1621 tagactggct tgaagcggat aaagttgcag gaccactact gcgttcggca cttcctgctg
     1681 gctggtttat tgctgataaa tctggggcag gagagcgtgg ttcacggggt atcattgccg
     1741 cacttggacc agatggtaag ccttcccgta tcgtagttat ctacacgacg ggtagtcagg
     1801 caactatgga cgaacgaaat agacagattg ctgaaatagg ggcttcactg attaagcatt
     1861 ggtaaaccga tacaattaaa ggctcctttt ggagcctttt tttttggacg gaccggtaga
     1921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2041 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2581 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2641 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2701 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2761 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     2821 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     2881 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2941 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3001 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3061 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3121 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     3181 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3241 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3301 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgtt
//
