pEarleyGate 301
Catalog No. PVT11165
Packing 2ug
pEarleyGate 301 Information
Function plant expression plasmid
Terminator: OCS Terminator
Plasmid tagging: C-HA
Prokaryotic resistance: kanamycin Kan
Screening marker: Bar
Clone strain: Escherichia coli DB3.1
Culture conditions: 37 C
pEarleyGate 301 Description
PEarleyGate 301 (C-HA) is a Gateway expression system for plant cells. pEarleyGate 301 allows for rapid recombinational cloning of DNA from Gateway entry vectors to be epitope tagged with HA tag at the C-terminus of the encoded protein. This vector allows for expression of genes from a promoter of choice rather than from the enhanced 35S promoter (see pEarleyGate 101-105 or 201-204).
pEarleyGate 301 Sequence
LOCUS Exported 10379 bp ds-DNA circular SYN 18-DEC-2017
DEFINITION Gateway(R)-compatible promoterless plant transformation vector with
a C-terminal HA tag.
KEYWORDS pEarleyGate 301
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10379)
AUTHORS Earley KW, Haag JR, Pontes O, Opper K, Juehne T, Song K, Pikaard CS.
TITLE Gateway-compatible vectors for plant functional genomics and
proteomics.
JOURNAL Plant J. 2006;45:616-29.
PUBMED 16441352
REFERENCE 2 (bases 1 to 10379)
AUTHORS Pikaard Lab
TITLE Direct Submission
COMMENT The sequence was corrected by inserting a G at position 6487.
FEATURES Location/Qualifiers
source 1..10379
/organism="synthetic DNA construct"
/lab_host="Plant Cells"
/mol_type="other DNA"
misc_feature 3..27
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
terminator 113..365
/label=MAS terminator
/note="mannopine synthase terminator"
CDS complement(375..926)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=Bar
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(932..1312)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
protein_bind 1364..1483
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1513..1543
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1597..2256
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=Chl
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2598..2903
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2944..3068)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 3070..3096
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
terminator 3150..3857
/label=OCS terminator
/note="octopine synthase terminator"
misc_feature 4101..4125
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 5425..6054
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 6483..7556
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 7622..7816
温馨提示：不可用于临床ZL。
