| 载体基本信息 |
| 载体名称 |
pDEST26 |
载体抗性 |
Ampicillin |
| 载体长度 |
7481 bp |
载体类型 |
Gateway Cloning Vectors |
| 载体来源 |
Invitrogen (Life Technologies) |
筛选标记 |
Neomycin/G418(Geneticin) |
| 拷贝数 |
High copy number |
5引物 |
M13 fwd |
| 3引物 |
M13 fwd |
pDEST26 载体载体图谱
pDEST26 载体载体序列
LOCUS Exported 7481 bp ds-DNA circular SYN 11-11-2013
DEFINITION Gateway(R) destination vector for high-level expression of
N-terminally 6xHis-tagged proteins in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pDEST26
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7481)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7481
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/mol_type="other DNA"
enhancer 15..318
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 319..522
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 632..634
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 644..661
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
protein_bind 671..795
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 820..850
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
CDS 904..1563
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1905..2210
/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(2251..2375)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(2408..2426)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2437..2453)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 2858..2992
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 3118..3573
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3692..4049
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 3900..4035
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 4100..4894
/codon_start=1
/gene="aph(3)-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 4958..5006
/note="synthetic polyadenylation signal"
promoter 5312..5416
/gene="bla"
/note="AmpR promoter"
CDS 5417..6277
/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 6448..7036
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"