LOCUS       Exported                4570 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pQEMBP, complete sequence.
ACCESSION   AY327141
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4570)
  AUTHORS   Binz HK, Amstutz P, Kohl A, Stumpp MT, Briand C, Forrer P, Grutter 
            MG, Pluckthun A.
  TITLE     High-affinity binders selected from designed ankyrin repeat protein 
            libraries
  JOURNAL   Nat. Biotechnol. 22 (5), 575-582 (2004)
  PUBMED    15097997
REFERENCE   2  (bases 1 to 4570)
  AUTHORS   Stumpp MT, Forrer P, Binz KH, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland
REFERENCE   3  (bases 1 to 4570)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4570
                     /organism="Expression vector pQEMBP"
                     /mol_type="genomic DNA"
                     /label=for the production of MBP and MBP fusion proteins
                     /note="for the production of MBP and MBP fusion proteins"
                     /db_xref="taxon:248465"
     regulatory      7..87
                     /regulatory_class="promoter"
                     /note="T5"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             115..1302
                     /codon_start=1
                     /product="MBPfusion"
                     /label=MBPfusion
                     /note="RGS-His-tag_MBP_mcs fusion protein; designed to 
                     increase ability to solubilize difficult proteins and 
                     improve expression yield"
                     /protein_id="AAQ93662.1"
                     /translation="MRGSHHHHHHGSGSMKTEEGKLVIWINGDKGYNGLAEVGKKFEKD
                     TGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKL
                     YPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALM
                     FNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNAD
                     TDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGI
                     NAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQ
                     KGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTGSGGTPGRPAAKLN"
     misc_feature    115..144
                     /label=RGS-His-tag
                     /note="RGS-His-tag"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    145..1257
                     /label=Escherichia coli maltose binding protein
                     /note="Escherichia coli maltose binding protein"
     CDS             157..1257
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     misc_feature    1258..1302
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      1312..1406
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2178..2264
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    2420..2560
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      2693..3349
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(2746..3334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3505..4365)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance"
                     /protein_id="AAQ93663.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4366..4470)
                     /gene="bla"
                     /label=AmpR promoter