LOCUS Exported 3781 bp ds-DNA circular SYN 23-11-2013
DEFINITION Gateway(R) Dual Selection Vector with two prokaryotic ribosome
binding sites for creating an entry vector by restriction cloning.
ACCESSION .
VERSION .
KEYWORDS pENTR11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3781)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
COMMENT To create a Kozak sequence, clone a blunt fragment starting with
ATGG (where ATG is the start codon) into the XmnI site.
FEATURES Location/Qualifiers
source 1..3781
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 103..189
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..457
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL1"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 457..526
/note="MCS 1"
/note="multiple cloning site, part 1"
RBS 461..466
/note="ribosome binding site"
RBS 483..488
/note="ribosome binding site"
CDS complement(567..872)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1192..1872)
/codon_start=1
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
promoter complement(1926..1956)
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
misc_feature 1981..2006
/note="MCS 2"
/note="multiple cloning site, part 2"
protein_bind complement(2010..2109)
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2232..3041
/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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 3131..3719
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 3781 bp ds-DNA circular SYN 23-11-2013
DEFINITION Gateway(R) Dual Selection Vector with two prokaryotic ribosome
binding sites for creating an entry vector by restriction cloning.
ACCESSION .
VERSION .
KEYWORDS pENTR11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3781)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
COMMENT To create a Kozak sequence, clone a blunt fragment starting with
ATGG (where ATG is the start codon) into the XmnI site.
FEATURES Location/Qualifiers
source 1..3781
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 103..189
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..457
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL1"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 457..526
/note="MCS 1"
/note="multiple cloning site, part 1"
RBS 461..466
/note="ribosome binding site"
RBS 483..488
/note="ribosome binding site"
CDS complement(567..872)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1192..1872)
/codon_start=1
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
promoter complement(1926..1956)
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
misc_feature 1981..2006
/note="MCS 2"
/note="multiple cloning site, part 2"
protein_bind complement(2010..2109)
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2232..3041
/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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 3131..3719
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"