LOCUS       Exported                3934 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKEx4tr, complete sequence.
ACCESSION   AF044029
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3934)
  AUTHORS   Mindrinos M, Katagiri F, Yu GL, Ausubel FM.
  TITLE     The A. thaliana disease resistance gene RPS2 encodes a protein 
            containing a nucleotide-binding site and leucine-rich repeats
  JOURNAL   Cell 78 (6), 1089-1099 (1994)
  PUBMED    7923358
REFERENCE   2  (bases 1 to 3934)
  AUTHORS   Katagiri F, Yuan F, Ausubel FM.
  TITLE     A plant cDNA expression vector carrying a modified version of the 
            cauliflower mosaic virus 35S promoter that has a reduced activity in
            bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3934)
  AUTHORS   Katagiri F, Ausubel FM.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JAN-1998) Biological Sciences, University of Maryland 
            Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA
REFERENCE   4  (bases 1 to 3934)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3934
                     /organism="Expression vector pKEx4tr"
                     /mol_type="genomic DNA"
                     /label=derived from pUC18 vector
                     /note="derived from pUC18 vector"
                     /db_xref="taxon:72436"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      417..1305
                     /regulatory_class="promoter"
                     /note="modified cauliflower mosaic virus 35S promoter"
     terminator      1199..1245
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      1312..1334
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        1312..1330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1335..1389
                     /note="multiple cloning site; contains unique restriction 
                     sites for PmeI, SalI, EcoRI, BamHI, XhoI, NotI, SacII, and 
                     SacI"
     regulatory      complement(1401..1420)
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        complement(1402..1420)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1437..1690
                     /regulatory_class="terminator"
                     /note="3 end of nopalin synthase gene"
     terminator      1437..1689
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1704..404
                     /label=similar to pUC18 vector
                     /note="similar to pUC18 vector"
     primer_bind     complement(1713..1729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1737..1753
                     /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)."
     promoter        complement(1761..1791)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1806..1827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2115..2703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2874..3734)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3735..3839)
                     /gene="bla"
                     /label=AmpR promoter