pEarleyGate 205 载体 (V010857#)

载体基本信息
载体名称 pEarleyGate 205 载体抗性 Chloramphenicol
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 20512188 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000LB T-DNA repeatMAS terminatorBlpRMAS promoterCaMV 35S promoterattR1lac UV5 promoterCmRccdBattR2CBPTEV SiteProtAProtAOCS terminatorRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanR

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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