| 载体基本信息 |
| 载体名称 |
pIPKb002 |
载体抗性 |
Spectinomycin/Streptomycin |
| 载体长度 |
13992 bp |
载体类型 |
Binary vector |
| 载体来源 |
Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer P, Kumlehn J. |
pIPKb002 载体载体图谱
pIPKb002 载体载体序列
LOCUS Exported 13992 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Binary vector pIPKb002, complete sequence.
ACCESSION EU161568
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13992)
AUTHORS Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer
P, Kumlehn J.
TITLE A set of modular binary vectors for transformation of cereals
JOURNAL Plant Physiol. 145 (4), 1192-1200 (2007)
PUBMED 17981986
REFERENCE 2 (bases 1 to 13992)
AUTHORS Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer
P, Kumlehn J.
TITLE Direct Submission
JOURNAL Submitted (18-SEP-2007) Cytogenetics and Molecular Cell Biology,
Leibniz Institute of Plant Genetics and Crop Plant Research,
Corrensstr. 3, Gatersleben D-06466, Germany
REFERENCE 3 (bases 1 to 13992)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13992
/organism="Binary vector pIPKb002"
/mol_type="other DNA"
/label=binary vector for over-expression in cereals
/note="binary vector for over-expression in cereals"
/db_xref="taxon:475864"
regulatory 10..996
/regulatory_class="promoter"
/note="maize ubiquitin (Ubi-1) promoter"
intron 997..1516
/note="maize ubiquitin (Ubi-1) intron 1"
CDS 1544..2569
/codon_start=1
/gene="hpt"
/product="hygromycin phosphotransferase"
/label=hpt
/protein_id="ABW70072.1"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
gene 1544..2569
/gene="hpt"
/label=hpt
regulatory 2594..2795
/regulatory_class="terminator"
/note="CaMV 35S terminator"
regulatory 2828..2837
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory complement(2847..3100)
/regulatory_class="terminator"
/note="nopaline synthase terminator"
terminator 2848..3100
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_recomb 3119..3243
/label=attR2
/note="attR2"
protein_bind 3119..3243
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(3284..3589)
/codon_start=1
/gene="ccdB"
/product="gyrase inhibitor"
/label=ccdB
/protein_id="ABW70070.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene complement(3284..3589)
/gene="ccdB"
/label=ccdB
CDS complement(3931..4590)
/codon_start=1
/gene="CmR"
/product="chloramphenicol resistance protein"
/label=CmR
/protein_id="ABW70071.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene complement(3931..4590)
/gene="CmR"
/label=CmR
promoter complement(4644..4674)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
misc_recomb 4699..4823
/label=attR1
/note="attR1"
protein_bind complement(4699..4823)
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
intron complement(4874..5393)
/note="maize ubiquitin (Ubi-1) intron 1"
regulatory complement(5394..6080)
/regulatory_class="promoter"
/note="maize ubiquitin (Ubi-1) promoter"
misc_feature 6466..6490
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
misc_feature 6467..6490
/label=right T-DNA border
/note="right T-DNA border"
rep_origin 7025..7722
/label=ColE1 origin of replication
/note="ColE1 origin of replication"
rep_origin 7135..7723
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 7908..8048
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 8271..11976
/label=pVS1 origin of replication
/note="pVS1 origin of replication"
CDS 9383..10012
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 10441..11514
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 11580..11774
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS 12160..13170
/codon_start=1
/gene="spec"
/product="streptomycin/spectinomycin adenyltransferase"
/label=spec
/protein_id="ABW70073.1"
/translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
gene 12160..13170
/gene="spec"
/label=spec
CDS 12379..13170
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 13690..13714
/label=LB T-DNA repeat
/note="left border repeat from octopine Ach5 T-DNA"
misc_feature 13691..13714
/label=left T-DNA border
/note="left T-DNA border"