LOCUS       Exported                3499 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pQStrep2, complete sequence.
ACCESSION   AY028642
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3499)
  AUTHORS   Scheich C, Sievert V, Bussow K.
  TITLE     An automated method for high-throughput protein purification applied
            to a comparison of His-tag and GST-tag affinity chromatography
  JOURNAL   BMC Biotechnol. 3, 12 (2003)
  PUBMED    12885298
REFERENCE   2  (bases 1 to 3499)
  AUTHORS   Bussow K, Tischer J.
  TITLE     Cloning vector pQStrep2
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3499)
  AUTHORS   Bussow K, Tischer J.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2001) Dept. Prof. H. Lehrach, Max Planck Institute
            of Molecular Genetics, Ihnestr. 73, Berlin 14195, Germany
REFERENCE   4  (bases 1 to 3499)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3499
                     /organism="Cloning vector pQStrep2"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:155767"
     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)."
     regulatory      30..35
                     /regulatory_class="minus_35_signal"
     regulatory      48..53
                     /regulatory_class="minus_10_signal"
     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)"
     regulatory      101..107
                     /regulatory_class="ribosome_binding_site"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    145..197
                     /note="multiple cloning site for BamHI, SalI, BglII and 
                     NotI"
     CDS             199..222
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      245..339
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             383..1042
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1107..1193
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1349..1489
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1675..2263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2434..3294)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AAK26193.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3295..3399)
                     /gene="bla"
                     /label=AmpR promoter