pEarleyGate 301

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  • Model: PVT11165
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pEarleyGate 301


Catalog No. PVT11165
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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
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