LOCUS       sequence-177525-        3409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 3 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 3409)
  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 3409)
  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-CBPHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3409
                     /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             215..232
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..280
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(841..859)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      855..902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        967..1058
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1923..1962
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1993..2581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2482..2501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2735..2752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2767..2906
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2832..2851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2992..3014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3008..3199)
                     /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 tagccatcat catcatcatc acagcagcgg
      241 atctggtggt ggtggtggtg aaaacctgta cttccaggga tccatcagtc tgattgcggc
      301 gctagcggta gatcgcgtta tcggcatgga aaacgccatg ccatggaacc tgcctgccga
      361 tctcgcctgg tttaaacgca acaccttaaa taaacccgtg attatggggc gccatacctg
      421 ggaatcaatc ggtaggcctt tgcccggccg caaaaatatt atcctcagca gtcaacccgg
      481 gaccgatgat cgggttacct gggttaaatc ggtcgacgaa gccatcgcgg cgtgtggtga
      541 cgtaccagaa atcatggtga ttggcggcgg acgcgtttat gaacagttct tgccaaaagc
      601 gcaaaaactg tatctgacgc atatcgatgc agaagtggaa ggcgacaccc attttccgga
      661 ttacgagccg gatgactggg aatcggtatt cagcgaattc cacgatgctg atgcgcagaa
      721 ctcgcatagc tattgtttcg aaatcctcga gcgtcgtgcc ggctaatgta caggtaccac
      781 gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      841 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      901 tgctgaaagg aggaactata tcgggattag cccgcctaat gagcgggctt ttttttaatt
      961 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1021 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1081 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1141 tcgtgaaagt aaaagacgca gaggaccaat tgggggcacg agtgggatac atagaactgg
     1201 acttgaatag cggtaaaatc cttgagagtt ttcgccctga agagcgtttt ccaatgatga
     1261 gcactttcaa agttctgcta tgtggagcag tattatcccg tgtagatgcg gggcaagagc
     1321 aactcggacg acgaatacac tattcgcaga atgacttggt tgaatactcc ccagtgacag
     1381 aaaagcacct tacggacgga atgacggtaa gagaattatg tagtgccgcc ataacgatga
     1441 gtgataacac tgcggcgaac ttacttctga caaccatcgg tggaccgaag gaattaaccg
     1501 cttttttgca caatatggga gaccatgtaa ctcgccttga ccgttgggaa ccagaactga
     1561 atgaagccat accaaacgac gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat
     1621 tacgcaaact attaactggc gaactactta ctctggcttc acggcaacaa ttaatagact
     1681 ggcttgaagc ggataaagtt gcaggaccac tactgcgttc ggcacttcct gctggctggt
     1741 ttattgctga taaatctggg gcaggagagc gtggttcacg gggtatcatt gccgcacttg
     1801 gaccagatgg taagccttcc cgtatcgtag ttatctacac gacgggtagt caggcaacta
     1861 tggacgaacg aaatagacag attgctgaaa taggggcttc actgattaag cattggtaaa
     1921 ccgatacaat taaaggctcc ttttggagcc tttttttttg gacggaccgg tagaaaagat
     1981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2101 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2341 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2401 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2461 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2521 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2581 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2641 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2701 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2761 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     2821 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2881 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2941 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3001 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3061 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3121 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3181 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atggggtaat
     3241 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     3301 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3361 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgtt
//
