LOCUS       sequence-177401-        3364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 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 3364)
  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 3364)
  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-CBPNDHFR
FEATURES             Location/Qualifiers
     source          1..3364
                     /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             221..241
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(796..814)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      810..857
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        922..1013
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1878..1917
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1948..2536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2437..2456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2690..2707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2722..2861
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2787..2806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2947..2969
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2963..3154)
                     /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 gatctggtgg tggtggtggt gaaaacctgt acttccaggg
      241 atccatcagt ctgattgcgg cgctagcggt agatcgcgtt atcggcatgg aaaacgccat
      301 gccatggaac ctgcctgccg atctcgcctg gtttaaacgc aacaccttaa ataaacccgt
      361 gattatgggg cgccatacct gggaatcaat cggtaggcct ttgcccggcc gcaaaaatat
      421 tatcctcagc agtcaacccg ggaccgatga tcgggttacc tgggttaaat cggtcgacga
      481 agccatcgcg gcgtgtggtg acgtaccaga aatcatggtg attggcggcg gacgcgttta
      541 tgaacagttc ttgccaaaag cgcaaaaact gtatctgacg catatcgatg cagaagtgga
      601 aggcgacacc cattttccgg attacgagcc ggatgactgg gaatcggtat tcagcgaatt
      661 ccacgatgct gatgcgcaga actcgcatag ctattgtttc gaaatcctcg agcgtcgtgc
      721 cggctaatgt acaacgcgtg cgcgctgatc cggctgctaa caaagcccga aaggaagctg
      781 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
      841 tcttgagggg ttttttgctg aaaggaggaa ctatatcggg attagcccgc ctaatgagcg
      901 ggcttttttt taattcccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1081 ctcacccaga aacgctcgtg aaagtaaaag acgcagagga ccaattgggg gcacgagtgg
     1141 gatacataga actggacttg aatagcggta aaatccttga gagttttcgc cctgaagagc
     1201 gttttccaat gatgagcact ttcaaagttc tgctatgtgg agcagtatta tcccgtgtag
     1261 atgcggggca agagcaactc ggacgacgaa tacactattc gcagaatgac ttggttgaat
     1321 actccccagt gacagaaaag caccttacgg acggaatgac ggtaagagaa ttatgtagtg
     1381 ccgccataac gatgagtgat aacactgcgg cgaacttact tctgacaacc atcggtggac
     1441 cgaaggaatt aaccgctttt ttgcacaata tgggagacca tgtaactcgc cttgaccgtt
     1501 gggaaccaga actgaatgaa gccataccaa acgacgagcg agacaccaca atgcctgcgg
     1561 caatggcaac aacattacgc aaactattaa ctggcgaact acttactctg gcttcacggc
     1621 aacaattaat agactggctt gaagcggata aagttgcagg accactactg cgttcggcac
     1681 ttcctgctgg ctggtttatt gctgataaat ctggggcagg agagcgtggt tcacggggta
     1741 tcattgccgc acttggacca gatggtaagc cttcccgtat cgtagttatc tacacgacgg
     1801 gtagtcaggc aactatggac gaacgaaata gacagattgc tgaaataggg gcttcactga
     1861 ttaagcattg gtaaaccgat acaattaaag gctccttttg gagccttttt ttttggacgg
     1921 accggtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1981 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2041 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2101 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2161 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2221 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2281 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2341 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2401 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2461 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2521 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2581 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2641 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2701 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2761 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2821 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     2881 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2941 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3001 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3061 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3121 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3181 tgttcatggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     3241 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     3301 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     3361 tgtt
//
