| 载体基本信息 |
| 载体名称 |
pGWB721 |
载体抗性 |
Spectinomycin/Streptomycin |
| 载体长度 |
12570 bp |
载体类型 |
Gateway binary vector |
| 载体来源 |
Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T. |
pGWB721 载体载体图谱
pGWB721 载体载体序列
LOCUS Exported 12570 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB721 DNA, complete sequence.
ACCESSION AB608287
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12570)
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 12570)
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 12570)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..12570
/organism="Gateway binary vector pGWB721"
/mol_type="other DNA"
/db_xref="taxon:944723"
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 1178..1582
/label=10xMyc
/note="10xMyc"
CDS 1193..1222
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1229..1258
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1274..1303
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1310..1339
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1346..1375
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1391..1420
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1427..1456
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1463..1492
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1508..1537
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1544..1573
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
misc_feature complement(1592..1716)
/label=attR1
/note="attR1"
protein_bind 1592..1716
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1753..1783
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1837..2496
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAJ61218.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1837..2496
/gene="cat"
/label=cat
CDS 2838..3143
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAJ61219.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2838..3143
/gene="ccdB"
/label=ccdB
misc_feature 3184..3308
/label=attR2
/note="attR2"
protein_bind complement(3184..3308)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3318..3727
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3334..3586
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3620..3636)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3644..3660
/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(3668..3698)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3713..3734
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
regulatory complement(3763..4022)
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3770..4022
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(4060..5355)
/codon_start=1
/gene="gpt"
/product="UDP-N-acetylglucosamine: dolichol phosphate
N-acetylglucosamine-1-P transferase"
/label=gpt
/protein_id="BAJ61220.1"
/translation="MAARKRASSISIPNKPDPSEPNSAPSEQKMTRKTVSASGEEFRLA
PPKLGVIFVISTLLCSLYLYLLCFHYKVDNELKRSILINAGLSLVGFFVTLKMIPVTAR
YVLRRNMFGFDINKRGTPQGDIKVPESLGIVVGIVFLIVAIIFQYFNFTEDSNWLVEYN
AALASICFMILLGFVDDVLDVPWRVKLVLPSFATLPLLMAYAGHTTIVIPKPLVAYIGL
EVLNLGRIYKLYMGLLAVFCTNSINIHAGLNGLEIGQTVVIAAAILIHNVMQIGASVDP
EYHQAHAFSIFLTQPLMATSLAMLAYNWYPSSVFVGDTYTVFAGMTMAVVGILGHFSET
LLIFFLPQVLNLLLSLPQLAGIVKCPRHRLPRYDPATGLLTGTKDGTLVNVYLRLFGPK
SEKSLCIHLLVFQALACAFCFILRHFLAGWYK"
gene complement(4060..5355)
/gene="gpt"
/label=gpt
regulatory complement(5383..5669)
/regulatory_class="promoter"
/note="NOS promoter"
promoter complement(5407..5586)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(6092..6116)
/label=T-DNA left border
/note="T-DNA left border"
CDS 6637..7428
/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 7674..8262
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 8448..8588
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 8932..9126
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9192..10265)
/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(10694..11323)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"