载体基本信息 | |||
载体名称 | pGWB460 | 载体抗性 | Spectinomycin/Streptomycin |
---|---|---|---|
载体长度 | 12399 bp | 载体类型 | Gateway binary vector |
载体来源 | Tanaka Y, Nakamura S, Nakagawa T. |
pGWB460 载体载体图谱
pGWB460 载体载体序列
LOCUS Exported 12399 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Gateway binary vector pGWB460 DNA, complete sequence. ACCESSION AB543174 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12399) AUTHORS Tanaka Y, Nakamura S, Nakagawa T. TITLE Development of the tagRFP Gateway Binary Vectors JOURNAL Published Only in Database (2010) REFERENCE 2 (bases 1 to 12399) AUTHORS Tanaka Y, Nakamura S, Nakagawa T. TITLE Direct Submission JOURNAL Submitted (22-JAN-2010) Contact:Tsuyoshi Nakagawa Shimane University, Center for Integrated Research in Science; 1060 Nishikawatsu, Matsue, Shimane 690-8504, Japan REFERENCE 3 (bases 1 to 12399) AUTHORS . TITLE Direct Submission COMMENT constructed using pPZP221. FEATURES Location/Qualifiers source 1..12399 /organism="Gateway binary vector pGWB460" /mol_type="other DNA" /db_xref="taxon:711458" 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="BAI67573.1" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" gene 1424..2083 /gene="cat" /label=cat CDS 2425..2730 /codon_start=1 /gene="ccdB" /label=ccdB /protein_id="BAI67574.1" /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" CDS 2906..3619 /codon_start=1 /gene="tagrfp" /product="tag red fluorescent protein" /label=tagrfp /protein_id="BAI67575.1" /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV ARYCDLPSKLGHKLN" gene 2906..3619 /gene="tagrfp" /label=tagrfp regulatory 3625..4034 /regulatory_class="terminator" /note="NOS terminator" terminator 3641..3893 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(3927..3943) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3951..3967 /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(3975..4005) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4020..4041 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." regulatory complement(4068..4327) /regulatory_class="terminator" /note="NOS terminator" terminator 4075..4327 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(4717..5511) /codon_start=1 /gene="nptII" /product="neomycin phosphotransferase II" /label=nptII /protein_id="BAI67576.1" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" gene complement(4717..5511) /gene="nptII" /label=nptII regulatory complement(5524..5830) /regulatory_class="promoter" /note="NOS promoter" promoter complement(5532..5715) /function="NOS promoter" /label=NOS promoter /note="nopaline synthase promoter" promoter complement(5568..5747) /label=NOS promoter /note="nopaline synthase promoter" misc_feature complement(5921..5945) /label=T-DNA left border /note="T-DNA left border" CDS 6466..7257 /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 7503..8091 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 8277..8417 /label=bom /note="basis of mobility region from pBR322" rep_origin 8761..8955 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(9021..10094) /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(10523..11152) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
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