| 载体基本信息 |
| 载体名称 |
pGWB720 |
载体抗性 |
Spectinomycin/Streptomycin |
| 载体长度 |
12565 bp |
载体类型 |
Gateway binary vector |
| 载体来源 |
Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T. |
pGWB720 载体载体图谱
pGWB720 载体载体序列
LOCUS Exported 12565 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB720 DNA, complete sequence.
ACCESSION AB608286
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12565)
AUTHORS Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T.
TITLE Development of a series of gateway binary vectors possessing a
tunicamycin resistance gene as a marker for the transformation of
Arabidopsis thaliana
JOURNAL Biosci. Biotechnol. Biochem. 75 (4), 804-807 (2011)
PUBMED 21512216
REFERENCE 2 (bases 1 to 12565)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (12-JAN-2011) Contact:Tsuyoshi Nakagawa Shimane
University, Center for Integrated Research in Science; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 12565)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..12565
/organism="Gateway binary vector pGWB720"
/mol_type="other DNA"
/db_xref="taxon:944722"
misc_feature complement(54..78)
/label=T-DNA right border
/note="T-DNA right border"
primer_bind 281..297
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 325..1159
/regulatory_class="promoter"
/note="35S promoter"
promoter 812..1157
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature complement(1179..1303)
/label=attR1
/note="attR1"
protein_bind 1179..1303
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1340..1370
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1424..2083
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAJ61215.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1424..2083
/gene="cat"
/label=cat
CDS 2425..2730
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAJ61216.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2425..2730
/gene="ccdB"
/label=ccdB
misc_feature 2771..2895
/label=attR2
/note="attR2"
protein_bind complement(2771..2895)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 2906..3307
/label=10xMyc
/note="10xMyc"
CDS 2921..2950
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2957..2986
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3002..3031
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3038..3067
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3074..3103
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3119..3148
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3155..3184
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3191..3220
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3236..3265
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3272..3301
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
regulatory 3313..3722
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3329..3581
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3615..3631)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3639..3655
/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(3663..3693)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3708..3729
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
regulatory complement(3758..4017)
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3765..4017
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(4055..5350)
/codon_start=1
/gene="gpt"
/product="UDP-N-acetylglucosamine: dolichol phosphate
N-acetylglucosamine-1-P transferase"
/label=gpt
/protein_id="BAJ61217.1"
/translation="MAARKRASSISIPNKPDPSEPNSAPSEQKMTRKTVSASGEEFRLA
PPKLGVIFVISTLLCSLYLYLLCFHYKVDNELKRSILINAGLSLVGFFVTLKMIPVTAR
YVLRRNMFGFDINKRGTPQGDIKVPESLGIVVGIVFLIVAIIFQYFNFTEDSNWLVEYN
AALASICFMILLGFVDDVLDVPWRVKLVLPSFATLPLLMAYAGHTTIVIPKPLVAYIGL
EVLNLGRIYKLYMGLLAVFCTNSINIHAGLNGLEIGQTVVIAAAILIHNVMQIGASVDP
EYHQAHAFSIFLTQPLMATSLAMLAYNWYPSSVFVGDTYTVFAGMTMAVVGILGHFSET
LLIFFLPQVLNLLLSLPQLAGIVKCPRHRLPRYDPATGLLTGTKDGTLVNVYLRLFGPK
SEKSLCIHLLVFQALACAFCFILRHFLAGWYK"
gene complement(4055..5350)
/gene="gpt"
/label=gpt
regulatory complement(5378..5664)
/regulatory_class="promoter"
/note="NOS promoter"
promoter complement(5402..5581)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(6087..6111)
/label=T-DNA left border
/note="T-DNA left border"
CDS 6632..7423
/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"
rep_origin 7669..8257
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 8443..8583
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 8927..9121
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9187..10260)
/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"
CDS complement(10689..11318)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"