LOCUS       Exported                4272 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCL476 DNA, complete sequence.
ACCESSION   AB248921
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4272)
  AUTHORS   Love CA, Lilley PE, Dixon NE.
  TITLE     Stable high-copy-number bacteriophage lambda promoter vectors for 
            overproduction of proteins in Escherichia coli
  JOURNAL   Gene 176 (1-2), 49-53 (1996)
  PUBMED    8918231
REFERENCE   2  (bases 1 to 4272)
  AUTHORS   Yasuda S.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-FEB-2006) Contact:Seiichi Yasuda National Institute of
            Genetics, Microbial Genetics; Yata 1111, Mishima, Shizuoka 411-8540,
            Japan
REFERENCE   3  (bases 1 to 4272)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4272
                     /organism="Cloning vector pCL476"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:370947"
     misc_feature    53..419
                     /label=partition(pSC101)
                     /note="partition(pSC101)"
     promoter        474..578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             579..1439
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BAE79435.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            579..1439
                     /gene="bla"
                     /label=bla
     rep_origin      1610..2198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2486..2507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      2522..2578
                     /regulatory_class="promoter"
                     /note="lac promoter operator region"
     promoter        2522..2552
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2560..2576
                     /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     2584..2600
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2856..3569)
                     /codon_start=1
                     /gene="CI857"
                     /product="temperature-sensitive lambda-repressor"
                     /label=CI857
                     /protein_id="BAE79436.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     gene            complement(2856..3569)
                     /gene="CI857"
                     /label=CI857
     regulatory      3617..3644
                     /regulatory_class="promoter"
                     /note="lambda PR"
     RBS             3658..3666
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     regulatory      3831..3859
                     /regulatory_class="promoter"
                     /note="lambda PL"
     misc_feature    3980..4107
                     /label=polylinker
                     /note="polylinker"
     RBS             3986..3994
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             4005..4022
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4115..4131)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"