载体基本信息 | |||
载体名称 | pN6-Bxb1 | 载体抗性 | Kanamycin |
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载体长度 | 12721 bp | 载体类型 | Binary vector |
载体来源 | Thomson JG, Thilmony R. |
pN6-Bxb1 载体载体图谱
pN6-Bxb1 载体载体序列
LOCUS Exported 12721 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Binary vector pN6-Bxb1, complete sequence. ACCESSION JQ305140 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12721) AUTHORS Thomson JG, Thilmony R. TITLE Direct Submission JOURNAL Submitted (19-DEC-2011) Crop Improvement and Utilization Research Unit, USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA REFERENCE 2 (bases 1 to 12721) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..12721 /organism="Binary vector pN6-Bxb1" /mol_type="other DNA" /db_xref="taxon:1167201" misc_feature complement(1..26) /label=Agrobacterium left border /note="Agrobacterium left border" misc_feature 1..25 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" regulatory complement(76..291) /regulatory_class="polyA_signal_sequence" /note="CaMV 35S polyA terminator" polyA_signal 103..277 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(317..1342) /codon_start=1 /gene="hptII" /product="hygromycin phosphotransferase II" /label=hptII /note="hygromycin resistance plant selectable marker" /protein_id="AFF61393.1" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK K" gene complement(317..1342) /gene="hptII" /label=hptII regulatory complement(1378..1820) /regulatory_class="promoter" /note="double enhanced CaMV 35S promoter" promoter complement(1410..2087) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" protein_bind 2278..2299 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2314..2344 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2352..2368 /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)." primer_bind 2376..2392 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 2388..2621 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC" misc_feature 2402..2458 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(2462..2478) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 2855..3200 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 3095..3125 /label=35S forward 'e' primer annealing site /note="35S forward 'e' primer annealing site" misc_recomb 3233..3284 /label=Bxb1 recombinase attP attachment site /note="Bxb1 recombinase attP attachment site" misc_feature 3285..4039 /label=stuffer DNA /note="stuffer DNA" CDS 3295..4014 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" misc_recomb 4040..4081 /label=Bxb1 recombinase attB attachment site /note="Bxb1 recombinase attB attachment site" CDS join(4089..4103,4294..6141) /codon_start=1 /gene="gusA" /product="beta-glucuronidase" /label=gusA /note="GUS reporter" /protein_id="AFF61394.1" /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASHHHHHHV" gene 4089..6141 /gene="gusA" /label=gusA CDS join(4089..4103,4294..6111) /codon_start=1 /product="beta-glucuronidase" /label=GUS /note="This version of the gusA gene has a 5' extension with a catalase intron to ensure expression in plants but not bacteria." /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ" intron 4104..4293 /gene="gusA" /note="catalase intron" primer_bind complement(4297..4322) /label=GUS reverse 'f' primer annealing site /note="GUS reverse 'f' primer annealing site" CDS 6118..6135 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 6170..6422 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" regulatory 6195..6425 /regulatory_class="polyA_signal_sequence" /note="nopaline synthase (nos) polyA terminator" misc_feature 6444..6468 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" misc_feature complement(6463..6488) /label=Agrobacterium right border /note="Agrobacterium right border" misc_feature 6489..12721 /label=pCAMBIA 1301 binary vector sequence /note="pCAMBIA 1301 binary vector sequence" CDS 7768..8397 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 8831..9898 /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 9964..10158 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 10502..10642 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(10828..11416) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(11503..12297) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
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