LOCUS       Exported                3494 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pQE30NST, complete sequence.
ACCESSION   AF074376
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3494)
  AUTHORS   Bussow K, Cahill D, Nietfeld W, Bancroft D, Scherzinger E, Lehrach 
            H, Walter G.
  TITLE     A method for global protein expression and antibody screening on 
            high-density filters of an arrayed cDNA library
  JOURNAL   Nucleic Acids Res. 26 (21), 5007-5008 (1998)
  PUBMED    9776767
REFERENCE   2  (bases 1 to 3494)
  AUTHORS   Bussow K, Cahill D, Nietfeld W, Bancroft D, Scherzinger E, Lehrach 
            H, Walter G, Doebeli H, Eggimann B, Gentz R, Hochuli E, Stueber D.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-1998) Lehrach, Max-Planck-Institut fuer Molekulare
            Genetik, Ihnestrasse 73, Berlin 14195, Germany
REFERENCE   3  (bases 1 to 3494)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3494
                     /organism="Cloning vector pQE30NST"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /note="derived from sequence deposited under GenBank 
                     Accession Number A02229"
                     /db_xref="taxon:78926"
     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"
     misc_feature    115
                     /note="translation start site for inserts cloned into the 
                     multiple cloning site"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    145..224
                     /note="multiple cloning site for BamHI, SalI, BglII, NotI 
                     and HindIII"
     regulatory      151..173
                     /regulatory_class="promoter"
                     /note="SP6 RNA polymerase promoter"
     promoter        152..170
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(199..218)
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     promoter        complement(201..219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      240..334
                     /regulatory_class="terminator"
                     /note="Lambda t0 transcriptional terminator"
     terminator      240..334
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             378..1037
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      1096..1193
                     /regulatory_class="terminator"
                     /note="rrnB T1 transcriptional terminator"
     terminator      1102..1188
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1344..1484
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1670..2258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2429..3289)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase precursor"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="penicillinase"
                     /protein_id="AAC73065.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2429..3289)
                     /gene="bla"
                     /label=bla
     promoter        complement(3290..3394)
                     /gene="bla"
                     /label=AmpR promoter