LOCUS Exported 4107 bp ds-DNA circular SYN 23-8-2014
DEFINITION Gateway(R) destination vector for generating an expression clone by
three-fragment multisite recombination.
ACCESSION .
VERSION .
KEYWORDS pDEST R4-R3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4107)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4107
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
primer_bind 1..17
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 36..160
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR4"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(201..506)
/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(848..1507)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(1561..1591)
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind complement(1617..1740)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR3"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1748..1764)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2238..2342
/gene="bla"
/note="AmpR promoter"
CDS 2343..3203
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3374..3962
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 4107 bp ds-DNA circular SYN 23-8-2014
DEFINITION Gateway(R) destination vector for generating an expression clone by
three-fragment multisite recombination.
ACCESSION .
VERSION .
KEYWORDS pDEST R4-R3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4107)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4107
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
primer_bind 1..17
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 36..160
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR4"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(201..506)
/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(848..1507)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(1561..1591)
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind complement(1617..1740)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR3"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1748..1764)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2238..2342
/gene="bla"
/note="AmpR promoter"
CDS 2343..3203
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3374..3962
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"