LOCUS       sequence-177383-        3376 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 3376)
  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 3376)
  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-CBPNDHFR
FEATURES             Location/Qualifiers
     source          1..3376
                     /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..202
                     /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             227..247
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(808..826)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      822..869
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        934..1025
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1890..1929
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1960..2548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2449..2468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2702..2719
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2734..2873
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2799..2818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2959..2981
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2975..3166)
                     /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 gtctaagcga cgatggaaaa agaatttcat agccgtctca gcagccaacc gctttaagaa
      181 aatctcatcc tccggggcac ttctgggatc tggtggtggt ggtggtgaaa acctgtactt
      241 ccagggatcc atcagtctga ttgcggcgct agcggtagat cgcgttatcg gcatggaaaa
      301 cgccatgcca tggaacctgc ctgccgatct cgcctggttt aaacgcaaca ccttaaataa
      361 acccgtgatt atggggcgcc atacctggga atcaatcggt aggcctttgc ccggccgcaa
      421 aaatattatc ctcagcagtc aacccgggac cgatgatcgg gttacctggg ttaaatcggt
      481 cgacgaagcc atcgcggcgt gtggtgacgt accagaaatc atggtgattg gcggcggacg
      541 cgtttatgaa cagttcttgc caaaagcgca aaaactgtat ctgacgcata tcgatgcaga
      601 agtggaaggc gacacccatt ttccggatta cgagccggat gactgggaat cggtattcag
      661 cgaattccac gatgctgatg cgcagaactc gcatagctat tgtttcgaaa tcctcgagcg
      721 tcgtgccggc taatgtacag gtaccacgcg tgcgcgctga tccggctgct aacaaagccc
      781 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      841 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcg ggattagccc
      901 gcctaatgag cgggcttttt tttaattccc ctatttgttt atttttctaa atacattcaa
      961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1081 ttcctgtttt tgctcaccca gaaacgctcg tgaaagtaaa agacgcagag gaccaattgg
     1141 gggcacgagt gggatacata gaactggact tgaatagcgg taaaatcctt gagagttttc
     1201 gccctgaaga gcgttttcca atgatgagca ctttcaaagt tctgctatgt ggagcagtat
     1261 tatcccgtgt agatgcgggg caagagcaac tcggacgacg aatacactat tcgcagaatg
     1321 acttggttga atactcccca gtgacagaaa agcaccttac ggacggaatg acggtaagag
     1381 aattatgtag tgccgccata acgatgagtg ataacactgc ggcgaactta cttctgacaa
     1441 ccatcggtgg accgaaggaa ttaaccgctt ttttgcacaa tatgggagac catgtaactc
     1501 gccttgaccg ttgggaacca gaactgaatg aagccatacc aaacgacgag cgagacacca
     1561 caatgcctgc ggcaatggca acaacattac gcaaactatt aactggcgaa ctacttactc
     1621 tggcttcacg gcaacaatta atagactggc ttgaagcgga taaagttgca ggaccactac
     1681 tgcgttcggc acttcctgct ggctggttta ttgctgataa atctggggca ggagagcgtg
     1741 gttcacgggg tatcattgcc gcacttggac cagatggtaa gccttcccgt atcgtagtta
     1801 tctacacgac gggtagtcag gcaactatgg acgaacgaaa tagacagatt gctgaaatag
     1861 gggcttcact gattaagcat tggtaaaccg atacaattaa aggctccttt tggagccttt
     1921 ttttttggac ggaccggtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1981 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2041 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2101 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2161 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2221 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2281 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2341 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2401 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2461 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2521 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2581 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2641 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2701 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2761 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     2821 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     2881 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2941 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3001 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3061 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     3121 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     3181 aagggggatt tctgttcatg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     3241 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     3301 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     3361 tacagatgta ggtgtt
//
