LOCUS       sequence-177494-        3427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 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 3427)
  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 3427)
  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-CBPHISNDHFR
FEATURES             Location/Qualifiers
     source          1..3427
                     /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             227..244
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             272..292
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(859..877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      873..920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        985..1076
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1941..1980
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2011..2599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2500..2519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2753..2770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2785..2924
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2850..2869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3010..3032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3026..3217)
                     /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 tctagccatc atcatcatca
      241 tcacagcagc ggatctggtg gtggtggtgg tgaaaacctg tacttccagg gatccatcag
      301 tctgattgcg gcgctagcgg tagatcgcgt tatcggcatg gaaaacgcca tgccatggaa
      361 cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg tgattatggg
      421 gcgccatacc tgggaatcaa tcggtaggcc tttgcccggc cgcaaaaata ttatcctcag
      481 cagtcaaccc gggaccgatg atcgggttac ctgggttaaa tcggtcgacg aagccatcgc
      541 ggcgtgtggt gacgtaccag aaatcatggt gattggcggc ggacgcgttt atgaacagtt
      601 cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgat gcagaagtgg aaggcgacac
      661 ccattttccg gattacgagc cggatgactg ggaatcggta ttcagcgaat tccacgatgc
      721 tgatgcgcag aactcgcata gctattgttt cgaaatcctc gagcgtcgtg ccggctaatg
      781 tacaaagctt ggtaccacgc gtgcgcgctg atccggctgc taacaaagcc cgaaaggaag
      841 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      901 gggtcttgag gggttttttg ctgaaaggag gaactatatc gggattagcc cgcctaatga
      961 gcgggctttt ttttaattcc cctatttgtt tatttttcta aatacattca aatatgtatc
     1021 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1081 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1141 ttgctcaccc agaaacgctc gtgaaagtaa aagacgcaga ggaccaattg ggggcacgag
     1201 tgggatacat agaactggac ttgaatagcg gtaaaatcct tgagagtttt cgccctgaag
     1261 agcgttttcc aatgatgagc actttcaaag ttctgctatg tggagcagta ttatcccgtg
     1321 tagatgcggg gcaagagcaa ctcggacgac gaatacacta ttcgcagaat gacttggttg
     1381 aatactcccc agtgacagaa aagcacctta cggacggaat gacggtaaga gaattatgta
     1441 gtgccgccat aacgatgagt gataacactg cggcgaactt acttctgaca accatcggtg
     1501 gaccgaagga attaaccgct tttttgcaca atatgggaga ccatgtaact cgccttgacc
     1561 gttgggaacc agaactgaat gaagccatac caaacgacga gcgagacacc acaatgcctg
     1621 cggcaatggc aacaacatta cgcaaactat taactggcga actacttact ctggcttcac
     1681 ggcaacaatt aatagactgg cttgaagcgg ataaagttgc aggaccacta ctgcgttcgg
     1741 cacttcctgc tggctggttt attgctgata aatctggggc aggagagcgt ggttcacggg
     1801 gtatcattgc cgcacttgga ccagatggta agccttcccg tatcgtagtt atctacacga
     1861 cgggtagtca ggcaactatg gacgaacgaa atagacagat tgctgaaata ggggcttcac
     1921 tgattaagca ttggtaaacc gatacaatta aaggctcctt ttggagcctt tttttttgga
     1981 cggaccggta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2101 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2641 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2701 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2761 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2821 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2881 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2941 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3001 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3061 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3121 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3181 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3241 ttctgttcat ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3301 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3361 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3421 aggtgtt
//
