| 载体基本信息 | |||
| 载体名称 | pN6-Bxb1 | 载体抗性 | Kanamycin |
|---|---|---|---|
| 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"
我们不提供该载体，载体信息仅供查询和参考。