| 载体基本信息 |
| 载体名称 |
pYL436 |
载体抗性 |
Spectinomycin/Streptomycin |
| 载体长度 |
12607 bp |
载体类型 |
C-terminal TAP T-DNA vector |
| 载体来源 |
Rubio V, Shen Y, Saijo Y, Liu Y, Gusmaroli G, Dinesh-Kumar SP, Deng XW. |
pYL436 载体载体图谱
pYL436 载体载体序列
LOCUS Exported 12607 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION C-terminal TAP T-DNA vector pYL436, complete sequence.
ACCESSION AY737283
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12607)
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 12607)
AUTHORS Liu Y, Dinesh-Kumar SP.
TITLE A C-terminal tandem affinity purification (TAP) T-DNA vector pYL436
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 12607)
AUTHORS Liu Y, Dinesh-Kumar SP.
TITLE Direct Submission
JOURNAL Submitted (28-AUG-2004) MCDB, Yale University, New Haven, CT 06520,
USA
REFERENCE 4 (bases 1 to 12607)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12607
/organism="C-terminal TAP T-DNA vector pYL436"
/mol_type="other DNA"
/label=derived from pPZP222
/note="derived from pPZP222"
/db_xref="taxon:293967"
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 845..2718
/label=attR1-CmR-ccdB-attR2 GATEWAY cassette
/note="attR1-CmR-ccdB-attR2 GATEWAY cassette"
protein_bind 845..969
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1143..1245
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 1246..1905
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2247..2552
/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
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2593..2717)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 2752..3009
/label=9x myc tag
/note="9x myc tag"
CDS 2755..2784
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2791..2820
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2827..2856
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2875..2904
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2911..2940
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2947..2976
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2995..3024
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3031..3060
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3067..3096
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
misc_feature 3163..3180
/label=6x HIS tag
/note="6x HIS tag"
CDS 3163..3180
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 3181..3204
/label=protease 3C cleavage site
/note="protease 3C cleavage site"
CDS 3181..3204
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
misc_feature 3244..3591
/label=2x IgG binding domain
/note="2x IgG binding domain"
CDS 3247..3417
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNDAQAPK"
CDS 3421..3591
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNDAQAPK"
misc_feature 3670..3932
/label=Nos terminator
/note="Nos terminator"
terminator 3675..3927
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3943..3959)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3967..3983
/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(3991..4021)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4036..4057
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 4222..4246
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 5546..6175
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 6609..7676
/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 7742..7936
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 8280..8420
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8606..9194)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9438..10229)
/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 10757..10781
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 10879..11053
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(11088..11621)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MVRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(11681..12359)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 12588..12604
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"