LOCUS Exported 6422 bp ds-DNA circular SYN 15-11-2013
DEFINITION Gateway(R) destination vector for inducible high-level expression of
proteins in bacterial cells.
ACCESSION .
VERSION .
KEYWORDS pDEST14
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6422)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6422
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 21..39
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 67..191
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 338..440
/note="cat promoter"
/note="promoter of the E. coli cat gene"
CDS 441..1100
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1442..1747
/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
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1788..1912)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
terminator 1979..2026
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 2533..2637
/gene="bla"
/note="AmpR promoter"
CDS 2638..3498
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3669..4257
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4687..4878)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
LOCUS Exported 6422 bp ds-DNA circular SYN 15-11-2013
DEFINITION Gateway(R) destination vector for inducible high-level expression of
proteins in bacterial cells.
ACCESSION .
VERSION .
KEYWORDS pDEST14
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6422)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6422
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 21..39
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 67..191
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 338..440
/note="cat promoter"
/note="promoter of the E. coli cat gene"
CDS 441..1100
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1442..1747
/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
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1788..1912)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
terminator 1979..2026
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 2533..2637
/gene="bla"
/note="AmpR promoter"
CDS 2638..3498
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3669..4257
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4687..4878)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"