LOCUS       sequence-177351-        3451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 1 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 3451)
  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 3451)
  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: pST50Trc1-CBPHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3451
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..226
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             239..256
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             284..304
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(883..901)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      897..944
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1009..1100
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1965..2004
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2035..2623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2524..2543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2777..2794
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2809..2948
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2874..2893)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3034..3056
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3050..3241)
                     /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 tccaattgga attcgctagc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatgtctaa gcgacgatgg aaaaagaatt tcatagccgt
      181 ctcagcagcc aaccgcttta agaaaatctc atcctccggg gcacttctgg gatctagcca
      241 tcatcatcat catcacagca gcggatctgg tggtggtggt ggtgaaaacc tgtacttcca
      301 gggatccatc agtctgattg cggcgctagc ggtagatcgc gttatcggca tggaaaacgc
      361 catgccatgg aacctgcctg ccgatctcgc ctggtttaaa cgcaacacct taaataaacc
      421 cgtgattatg gggcgccata cctgggaatc aatcggtagg cctttgcccg gccgcaaaaa
      481 tattatcctc agcagtcaac ccgggaccga tgatcgggtt acctgggtta aatcggtcga
      541 cgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg gtgattggcg gcggacgcgt
      601 ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg acgcatatcg atgcagaagt
      661 ggaaggcgac acccattttc cggattacga gccggatgac tgggaatcgg tattcagcga
      721 attccacgat gctgatgcgc agaactcgca tagctattgt ttcgaaatcc tcgagcgtcg
      781 tgccggctaa tgtacaagat ccagatctaa gcttggtacc acgcgtgcgc gctgatccgg
      841 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
      901 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
      961 tatcgggatt agcccgccta atgagcgggc ttttttttaa ttcccctatt tgtttatttt
     1021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1081 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1141 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg
     1201 cagaggacca attgggggca cgagtgggat acatagaact ggacttgaat agcggtaaaa
     1261 tccttgagag ttttcgccct gaagagcgtt ttccaatgat gagcactttc aaagttctgc
     1321 tatgtggagc agtattatcc cgtgtagatg cggggcaaga gcaactcgga cgacgaatac
     1381 actattcgca gaatgacttg gttgaatact ccccagtgac agaaaagcac cttacggacg
     1441 gaatgacggt aagagaatta tgtagtgccg ccataacgat gagtgataac actgcggcga
     1501 acttacttct gacaaccatc ggtggaccga aggaattaac cgcttttttg cacaatatgg
     1561 gagaccatgt aactcgcctt gaccgttggg aaccagaact gaatgaagcc ataccaaacg
     1621 acgagcgaga caccacaatg cctgcggcaa tggcaacaac attacgcaaa ctattaactg
     1681 gcgaactact tactctggct tcacggcaac aattaataga ctggcttgaa gcggataaag
     1741 ttgcaggacc actactgcgt tcggcacttc ctgctggctg gtttattgct gataaatctg
     1801 gggcaggaga gcgtggttca cggggtatca ttgccgcact tggaccagat ggtaagcctt
     1861 cccgtatcgt agttatctac acgacgggta gtcaggcaac tatggacgaa cgaaatagac
     1921 agattgctga aataggggct tcactgatta agcattggta aaccgataca attaaaggct
     1981 ccttttggag cctttttttt tggacggacc ggtagaaaag atcaaaggat cttcttgaga
     2041 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2101 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2161 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2221 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2281 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2341 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2401 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2461 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2521 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2581 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2641 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2701 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2761 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     2821 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     2881 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2941 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3001 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3061 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3121 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     3181 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     3241 ctgatgcctc cgtgtaaggg ggatttctgt tcatggggta atgataccga tgaaacgaga
     3301 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     3361 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     3421 ccagcgcttc gttaatacag atgtaggtgt t
//
