载体基本信息 | |||
载体名称 | pCLEAN-G144 | 载体长度 | 4924 bp |
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载体类型 | Binary vector | 载体来源 | Thole V, Worland B, Snape JW, Vain P. |
pCLEAN-G144 载体载体图谱
pCLEAN-G144 载体载体序列
LOCUS Exported 4924 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Binary vector pCLEAN-G144, complete sequence. ACCESSION EU186089 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4924) AUTHORS Thole V, Worland B, Snape JW, Vain P. TITLE The pCLEAN dual binary vector system for Agrobacterium-mediated plant transformation JOURNAL Plant Physiol. 145 (4), 1211-1219 (2007) PUBMED 17932303 REFERENCE 2 (bases 1 to 4924) AUTHORS Thole V, Vain P. TITLE Direct Submission JOURNAL Submitted (01-OCT-2007) Department of Crop Genetics, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK REFERENCE 3 (bases 1 to 4924) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..4924 /organism="Binary vector pCLEAN-G144" /mol_type="other DNA" /db_xref="taxon:478690" misc_feature 1..1734 /note="Cloning vector pGreenII0000 (Genbank Accession Number EF590266)-based backbone sequence" rep_origin 1..720 /label=pUC-ori /note="pUC-ori" rep_origin complement(61..649) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(820..1635) /codon_start=1 /gene="nptI" /product="NptI" /label=nptI /note="kanamycin resistance" /protein_id="ABW84179.1" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" gene complement(820..1635) /gene="nptI" /label=nptI misc_feature complement(1774..3336) /note="CaMV35S:mGFP5-ER:SPA expression unit inserted in vector backbone sequence" regulatory complement(1774..2070) /regulatory_class="terminator" /note="soybean-derived poly(A)signal" CDS complement(2091..2882) /codon_start=1 /gene="mGFP5-ER" /product="mGFP5-ER" /label=mGFP5-ER /note="modified green fluorescent protein mGFP5-ER gene (Haseloff et al. (1997) Proc Natl Acad Sci USA 94: 2122-2127; Genbank Accession Number U87974) containing one nucleotide variation resulting in an amino acid change at position 46 (His[GTG] to Tyr [GTA])" /protein_id="ABW84180.1" /translation="MKTNLFLFLIFSLLLSLSSAEFSKGEELFTGVVPILVELDGDVNG YKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDF FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE YNYNSHNVYIMADKQKNGIKANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYL STQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKHDEL" gene complement(2091..2882) /gene="mGFP5-ER" /label=mGFP5-ER CDS complement(2106..2816) /codon_start=1 /gene="mgfp5" /product="GFP with folding enhancement mutations" /label=mgfp5 /note="suitable for expression in plants due to removal of a cryptic intron" /translation="SKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" regulatory complement(2909..3336) /regulatory_class="promoter" /note="35S promoter of CaMV" promoter complement(2915..3259) /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" misc_feature 3368..4117 /note="Cloning vector pGreenII0000 (Genbank Accession Number EF590266)-based backbone sequence" rep_origin 3551..4103 /label=pSa-ori /note="pSa-ori" rep_origin 3556..3991 /label=pSa ori /note="origin of replication from bacterial plasmid pSa" misc_feature 4118..4894 /note="T-DNA region based on cloning vector pGreenII0000 T-DNA region (Genbank Accession Number EU048862; Hellens et al. (2000) Plant Mol Biol 42: 819-832)" misc_feature 4118..4141 /note="24-nt long nopaline-type T-DNA left border sequence derived from T-DNA region (Genbank Accession Number EU048862; Hellens et al. (2000) Plant Mol Biol 42: 819-832) of cloning vector pGreenII0000; this left border sequence is one nucleotide shorter than the consensus sequence (Slightom et al., (1985) EMBO J 4:3069-3077)" misc_feature 4118..4140 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA (truncated)" CDS complement(4156..4521) /codon_start=1 /gene="lacZ" /product="LacZ" /label=lacZ /protein_id="ABW84181.1" /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR TDRPSQQLRSLNGEWKL" gene complement(4156..4521) /gene="lacZ" /label=lacZ primer_bind 4305..4321 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 4331..4349 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 4358..4465 /label=multiple cloning site /note="multiple cloning site" primer_bind 4375..4391 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4425..4441) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(4478..4496) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4517..4533) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4541..4557 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4565..4595) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4610..4631 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_feature 4870..4894 /note="25-nt long consensus T-DNA right border sequence of the nopaline vector pTiT37 (Slightom et al., (1985) EMBO J 4:3069-3077)" misc_feature 4895..4918 /note="right border overdrive sequence (Peralta et al. (1986) EMBO J 5: 1137-1142)"
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