载体基本信息 | |||
载体名称 | pEarleyGate 205 | 载体抗性 | Chloramphenicol |
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载体长度 | 12188 bp | 载体类型 | Plant Vectors |
载体来源 | Earley KW, Haag JR, Pontes O, Opper K, Juehne T, Song K, Pikaard CS. | 拷贝数 | High copy number |
pEarleyGate 205 载体载体图谱
pEarleyGate 205 载体载体序列
LOCUS Exported 12188 bp ds-DNA circular SYN 11-1-2016 DEFINITION Gateway(R)-compatible plant transformation vector with a C-terminal TAP tag. ACCESSION . VERSION . KEYWORDS pEarleyGate 205 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12188) 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 12188) AUTHORS Pikaard Lab TITLE Direct Submission COMMENT The sequence was corrected by inserting a G at position 8296. FEATURES Location/Qualifiers source 1..12188 /organism="synthetic DNA construct" /lab_host="Plant Cells" /mol_type="other DNA" misc_feature 1..25 /note="LB T-DNA repeat" /note="left border repeat from nopaline C58 T-DNA" terminator 111..363 /note="MAS terminator" /note="mannopine synthase terminator" CDS complement(373..924) /codon_start=1 /gene="Streptomyces hygroscopicus bar" /product="phosphinothricin acetyltransferase" /function=" " /note="BlpR" /note="confers resistance to bialophos or phosphinothricin" /protein_id=" " /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF WQLDFSLPVPPRPVLPVTEI" promoter complement(930..1310) /note="MAS promoter" /note="mannopine synthase promoter (Velten et al., 1984)" promoter 2306..2651 /note="CaMV 35S promoter" /note="strong constitutive promoter from cauliflower mosaic virus" protein_bind 2663..2782 /gene="mutant version of attR" /bound_moiety="LR Clonase(TM)" /note="attR1" /note="recombination site for the Gateway(R) LR reaction" promoter 2812..2842 /note="lac UV5 promoter" /note="E. coli lac promoter with an ""up"" mutation" CDS 2896..3555 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 3897..4202 /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(4243..4367) /gene="mutant version of attR" /bound_moiety="LR Clonase(TM)" /note="attR2" /note="recombination site for the Gateway(R) LR reaction" CDS 4381..4458 /codon_start=1 /product="calmodulin-binding peptide" /note="CBP" /note="derived from skeletal muscle myosin light chain kinase; binds calmodulin with nanomolar affinity in the presence of calcium" /translation="KRRWKKNFIAVSAANRFKKISSSGAL" CDS 4486..4506 /codon_start=1 /product="tobacco etch virus (TEV) recognition and cleavage site" /note="TEV Site" /translation="ENLYFQG" CDS 4546..4719 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /note="ProtA" /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL AEAKKLNDAQAPK" CDS 4720..4893 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /note="ProtA" /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL AEAKKLNGAQAPK" terminator 4959..5666 /note="OCS terminator" /note="octopine synthase terminator" misc_feature 5910..5934 /note="RB T-DNA repeat" /note="right border repeat from nopaline C58 T-DNA" CDS 7234..7863 /codon_start=1 /product="stability protein from Pseudomonas plasmid pVS1" /note="pVS1 StaA" /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 8292..9365 /codon_start=1 /product="replication protein from Pseudomonas plasmid pVS1" /note="pVS1 RepA" /note=" " /protein_id=" " /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 9431..9625 /note="pVS1 oriV" /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 9969..10109 /note="bom" /note="basis of mobility region from pBR322" rep_origin complement(10295..10883) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(10970..11764) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin" /protein_id=" " /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
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