LOCUS Exported 2275 bp ds-DNA circular SYN 07-1-2015
DEFINITION Vector for Expresso(R) cloning to add a C-terminal 6xHis tag, with
rhamnose-inducible expression.
ACCESSION .
VERSION .
KEYWORDS pRham C-His Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2275)
AUTHORS Lucigen
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2275
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 1..119
/note="rhaB promoter"
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
RBS 153..175
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 183..185
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 186..203
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 235..282
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 380..409
/note="T3Te terminator"
/note="phage T3 early transcription terminator"
rep_origin 431..1018
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 1030..1057
/note="T7Te terminator"
/note="phage T7 early transcription terminator"
CDS complement(1081..1272)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1273..2088)
/codon_start=1
/gene="aph(3)-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter complement(2089..2179)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
terminator 2180..2211
/note="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"
LOCUS Exported 2275 bp ds-DNA circular SYN 07-1-2015
DEFINITION Vector for Expresso(R) cloning to add a C-terminal 6xHis tag, with
rhamnose-inducible expression.
ACCESSION .
VERSION .
KEYWORDS pRham C-His Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2275)
AUTHORS Lucigen
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2275
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 1..119
/note="rhaB promoter"
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
RBS 153..175
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 183..185
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 186..203
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 235..282
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 380..409
/note="T3Te terminator"
/note="phage T3 early transcription terminator"
rep_origin 431..1018
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 1030..1057
/note="T7Te terminator"
/note="phage T7 early transcription terminator"
CDS complement(1081..1272)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1273..2088)
/codon_start=1
/gene="aph(3)-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter complement(2089..2179)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
terminator 2180..2211
/note="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"