载体基本信息 | |||
载体名称 | pN-TAPa | 载体抗性 | Spectinomycin/Streptomycin |
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载体长度 | 12537 bp | 载体类型 | N-terminal TAPa T-DNA vector |
载体来源 | Rubio V, Shen Y, Saijo Y, Liu Y, Gusmaroli G, Dinesh-Kumar SP, Deng XW. |
pN-TAPa 载体载体图谱
pN-TAPa 载体载体序列
LOCUS Exported 12537 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION N-terminal TAPa T-DNA vector pN-TAPa, complete sequence. ACCESSION AY788908 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12537) AUTHORS Rubio V, Shen Y, Saijo Y, Liu Y, Gusmaroli G, Dinesh-Kumar SP, Deng XW. TITLE An alternative tandem affinity purification strategy applied to Arabidopsis protein complex isolation JOURNAL Plant J. 41 (5), 767-778 (2005) PUBMED 15703063 REFERENCE 2 (bases 1 to 12537) AUTHORS Rubio V, Deng XW. TITLE Direct Submission JOURNAL Submitted (21-OCT-2004) MCDB, Yale University, 165, Prospect St., New Haven, CT 06511, USA REFERENCE 3 (bases 1 to 12537) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..12537 /organism="N-terminal TAPa T-DNA vector pN-TAPa" /mol_type="other DNA" /label=derived from pPZP222 /note="derived from pPZP222" /db_xref="taxon:300483" regulatory 1..769 /regulatory_class="promoter" /note="duplicated CaMV 35S promoter" promoter 724..769 /label=minimal CaMV 35S promoter /note="minimal 35S promoter from cauliflower mosaic virus" misc_feature 780..844 /label=TMV U1 omega sequence /note="TMV U1 omega sequence" misc_feature 791..846 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" misc_feature 854..1231 /label=2x protein A IgG binding domain /note="2x protein A IgG binding domain" regulatory 857..866 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 887..1057 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNDAQAPK" CDS 1061..1231 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNDAQAPK" misc_feature 1256..1279 /label=protease 3C cleavage site /note="protease 3C cleavage site" CDS 1256..1279 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" misc_feature 1286..1303 /label=6x HIS tag /note="6x HIS tag" CDS 1286..1303 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" misc_feature 1325..1675 /label=9x myc tag /note="9x myc tag" CDS 1328..1357 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1364..1393 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1400..1429 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1448..1477 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1484..1513 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1520..1549 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1568..1597 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1604..1633 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1640..1669 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" misc_feature 1704..3577 /label=attR1-CmR-ccdB-attR2 GATEWAY cassette /note="attR1-CmR-ccdB-attR2 GATEWAY cassette" protein_bind 1704..1828 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 2002..2104 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 2105..2764 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 3106..3411 /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 VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(3452..3576) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" terminator 3605..3857 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" regulatory 3607..3863 /regulatory_class="terminator" /note="NOS terminator" primer_bind complement(3873..3889) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3897..3913 /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(3921..3951) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3966..3987 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_feature 4152..4176 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 5476..6105 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 6539..7606 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" rep_origin 7672..7866 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 8210..8350 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(8536..9124) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(9368..10159) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" misc_feature 10687..10711 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal 10809..10983 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(11018..11551) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MVRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(11611..12289) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" primer_bind 12518..12534 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants"
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