LOCUS       Exported                4750 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRKW2, complete sequence.
ACCESSION   DQ657243
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4750)
  AUTHORS   Wu W, Niehrs C.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2006) Division of Molecular Embryology (A050), 
            Deutsches Krebsforschungszetrum (DKFZ), Im Neuenheimer Feld 280, 
            Heidelberg 69120, Germany
REFERENCE   2  (bases 1 to 4750)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4750
                     /organism="Cloning vector pRKW2"
                     /mol_type="other DNA"
                     /db_xref="taxon:387935"
     regulatory      1..618
                     /regulatory_class="promoter"
                     /note="CMV"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     822..845
                     /label=forward primer AG195
                     /note="forward primer AG195"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    949..980
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(956..972)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1008..1142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(1008..1135)
                     /label=SV40 polyA
                     /note="SV40 polyA"
     primer_bind     complement(1051..1073)
                     /label=reverse primer AG189
                     /note="reverse primer AG189"
     promoter        1215..1572
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      1234..1577
                     /regulatory_class="promoter"
                     /note="SV40"
     rep_origin      1423..1558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1592..1608)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1821..2276
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2608..2712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2713..3573
                     /codon_start=1
                     /gene="Amp"
                     /product="beta-lactamase"
                     /label=Amp
                     /protein_id="ABF81660.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2713..3573
                     /gene="Amp"
                     /label=Amp
     rep_origin      3744..4332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4656..4686
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4694..4710
                     /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)."
     primer_bind     4718..4734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"