LOCUS       sequence-177553-        3397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3397)
  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 3397)
  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-CBPHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3397
                     /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             119..196
                     /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             209..226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             254..274
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(829..847)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      843..890
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        955..1046
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1911..1950
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1981..2569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2470..2489
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2723..2740
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2755..2894
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2820..2839)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2980..3002
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2996..3187)
                     /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 atatgtctaa
      121 gcgacgatgg aaaaagaatt tcatagccgt ctcagcagcc aaccgcttta agaaaatctc
      181 atcctccggg gcacttctgg gatctagcca tcatcatcat catcacagca gcggatctgg
      241 tggtggtggt ggtgaaaacc tgtacttcca gggatccatc agtctgattg cggcgctagc
      301 ggtagatcgc gttatcggca tggaaaacgc catgccatgg aacctgcctg ccgatctcgc
      361 ctggtttaaa cgcaacacct taaataaacc cgtgattatg gggcgccata cctgggaatc
      421 aatcggtagg cctttgcccg gccgcaaaaa tattatcctc agcagtcaac ccgggaccga
      481 tgatcgggtt acctgggtta aatcggtcga cgaagccatc gcggcgtgtg gtgacgtacc
      541 agaaatcatg gtgattggcg gcggacgcgt ttatgaacag ttcttgccaa aagcgcaaaa
      601 actgtatctg acgcatatcg atgcagaagt ggaaggcgac acccattttc cggattacga
      661 gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc agaactcgca
      721 tagctattgt ttcgaaatcc tcgagcgtcg tgccggctaa tgtacaacgc gtgcgcgctg
      781 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      841 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      901 gaactatatc gggattagcc cgcctaatga gcgggctttt ttttaattcc cctatttgtt
      961 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1021 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1081 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctc gtgaaagtaa
     1141 aagacgcaga ggaccaattg ggggcacgag tgggatacat agaactggac ttgaatagcg
     1201 gtaaaatcct tgagagtttt cgccctgaag agcgttttcc aatgatgagc actttcaaag
     1261 ttctgctatg tggagcagta ttatcccgtg tagatgcggg gcaagagcaa ctcggacgac
     1321 gaatacacta ttcgcagaat gacttggttg aatactcccc agtgacagaa aagcacctta
     1381 cggacggaat gacggtaaga gaattatgta gtgccgccat aacgatgagt gataacactg
     1441 cggcgaactt acttctgaca accatcggtg gaccgaagga attaaccgct tttttgcaca
     1501 atatgggaga ccatgtaact cgccttgacc gttgggaacc agaactgaat gaagccatac
     1561 caaacgacga gcgagacacc acaatgcctg cggcaatggc aacaacatta cgcaaactat
     1621 taactggcga actacttact ctggcttcac ggcaacaatt aatagactgg cttgaagcgg
     1681 ataaagttgc aggaccacta ctgcgttcgg cacttcctgc tggctggttt attgctgata
     1741 aatctggggc aggagagcgt ggttcacggg gtatcattgc cgcacttgga ccagatggta
     1801 agccttcccg tatcgtagtt atctacacga cgggtagtca ggcaactatg gacgaacgaa
     1861 atagacagat tgctgaaata ggggcttcac tgattaagca ttggtaaacc gatacaatta
     1921 aaggctcctt ttggagcctt tttttttgga cggaccggta gaaaagatca aaggatcttc
     1981 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2041 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2101 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2161 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2221 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2281 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2341 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2401 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2461 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2521 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2581 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2641 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2701 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     2761 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     2821 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2881 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2941 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3001 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3061 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3121 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3181 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     3241 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3301 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3361 tcaatgccag cgcttcgtta atacagatgt aggtgtt
//
