LOCUS Exported 2235 bp ds-DNA circular SYN 08-1-2015
DEFINITION Vector for Expresso(R) cloning to add an N-terminal 6xHis tag, with
inducible expression from the T7 promoter.
ACCESSION .
VERSION .
KEYWORDS pETite N-His Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2235)
AUTHORS Lucigen
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2235
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 51..69
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 71..86
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 113..135
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 143..145
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 146..163
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 195..242
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 340..369
/note="T3Te terminator"
/note="phage T3 early transcription terminator"
rep_origin 391..978
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 990..1017
/note="T7Te terminator"
/note="phage T7 early transcription terminator"
CDS complement(1041..1232)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1233..2048)
/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(2049..2139)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
terminator 2140..2171
/note="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"
LOCUS Exported 2235 bp ds-DNA circular SYN 08-1-2015
DEFINITION Vector for Expresso(R) cloning to add an N-terminal 6xHis tag, with
inducible expression from the T7 promoter.
ACCESSION .
VERSION .
KEYWORDS pETite N-His Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2235)
AUTHORS Lucigen
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2235
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 51..69
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 71..86
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 113..135
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 143..145
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 146..163
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 195..242
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 340..369
/note="T3Te terminator"
/note="phage T3 early transcription terminator"
rep_origin 391..978
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 990..1017
/note="T7Te terminator"
/note="phage T7 early transcription terminator"
CDS complement(1041..1232)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1233..2048)
/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(2049..2139)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
terminator 2140..2171
/note="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"