LOCUS       sequence-177365-        3394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 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 3394)
  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 3394)
  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: pST50Trc2-CBPNDHFR
FEATURES             Location/Qualifiers
     source          1..3394
                     /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             101..123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             137..214
                     /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..259
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(826..844)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      840..887
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        952..1043
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1908..1947
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1978..2566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2467..2486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2720..2737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2752..2891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2817..2836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2977..2999
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2993..3184)
                     /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 attcaataat tttgtttaac tttaagaagg
      121 agatatacat atgtctaagc gacgatggaa aaagaatttc atagccgtct cagcagccaa
      181 ccgctttaag aaaatctcat cctccggggc acttctggga tctggtggtg gtggtggtga
      241 aaacctgtac ttccagggat ccatcagtct gattgcggcg ctagcggtag atcgcgttat
      301 cggcatggaa aacgccatgc catggaacct gcctgccgat ctcgcctggt ttaaacgcaa
      361 caccttaaat aaacccgtga ttatggggcg ccatacctgg gaatcaatcg gtaggccttt
      421 gcccggccgc aaaaatatta tcctcagcag tcaacccggg accgatgatc gggttacctg
      481 ggttaaatcg gtcgacgaag ccatcgcggc gtgtggtgac gtaccagaaa tcatggtgat
      541 tggcggcgga cgcgtttatg aacagttctt gccaaaagcg caaaaactgt atctgacgca
      601 tatcgatgca gaagtggaag gcgacaccca ttttccggat tacgagccgg atgactggga
      661 atcggtattc agcgaattcc acgatgctga tgcgcagaac tcgcatagct attgtttcga
      721 aatcctcgag cgtcgtgccg gctaatgtac aaagcttggt accacgcgtg cgcgctgatc
      781 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
      841 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
      901 ctatatcggg attagcccgc ctaatgagcg ggcttttttt taattcccct atttgtttat
      961 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1021 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1081 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctcgtg aaagtaaaag
     1141 acgcagagga ccaattgggg gcacgagtgg gatacataga actggacttg aatagcggta
     1201 aaatccttga gagttttcgc cctgaagagc gttttccaat gatgagcact ttcaaagttc
     1261 tgctatgtgg agcagtatta tcccgtgtag atgcggggca agagcaactc ggacgacgaa
     1321 tacactattc gcagaatgac ttggttgaat actccccagt gacagaaaag caccttacgg
     1381 acggaatgac ggtaagagaa ttatgtagtg ccgccataac gatgagtgat aacactgcgg
     1441 cgaacttact tctgacaacc atcggtggac cgaaggaatt aaccgctttt ttgcacaata
     1501 tgggagacca tgtaactcgc cttgaccgtt gggaaccaga actgaatgaa gccataccaa
     1561 acgacgagcg agacaccaca atgcctgcgg caatggcaac aacattacgc aaactattaa
     1621 ctggcgaact acttactctg gcttcacggc aacaattaat agactggctt gaagcggata
     1681 aagttgcagg accactactg cgttcggcac ttcctgctgg ctggtttatt gctgataaat
     1741 ctggggcagg agagcgtggt tcacggggta tcattgccgc acttggacca gatggtaagc
     1801 cttcccgtat cgtagttatc tacacgacgg gtagtcaggc aactatggac gaacgaaata
     1861 gacagattgc tgaaataggg gcttcactga ttaagcattg gtaaaccgat acaattaaag
     1921 gctccttttg gagccttttt ttttggacgg accggtagaa aagatcaaag gatcttcttg
     1981 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2041 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2101 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2161 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2221 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2281 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2341 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2401 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2461 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2521 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2581 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2641 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2701 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2761 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     2821 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     2881 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2941 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3001 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3061 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3121 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3181 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg gtaatgatac cgatgaaacg
     3241 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     3301 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     3361 atgccagcgc ttcgttaata cagatgtagg tgtt
//
