pGWB702omega 载体图谱和序列

我们所展示的质粒图谱主要是从文献和开放数据库中收集而来，主要是为了方便研究工作，其中一小部分质粒进行了质量控制，可供科学家使用。

所有的产品都严格仅供科学研究使用，不能应用于临床试验，包括人体摄入、注射或外用的药理学用途。

确保质粒的关键元件正确，但是我们并不能保证实验结果。页面展示的图谱序列为理论序列，可能与测序结果不一致，请自行比对后确定是否满足要求。(如果测过序，本页面一般会提供下载)

开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%，则将其视为正确。

由于科学研究是在探索未知，具有很大的不确定性，在任何情况下，我们都不承担超出质粒本身的额外经济损失或责任。

载体名称:: pGWB702omega

载体抗性:: Streptomycin

载体长度:: 12156 bp

载体类型:: Gateway binary vector

复制子:: ori

载体来源:: Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T.

启动子:: CaMV35S(enhanced)

pGWB702omega 载体图谱

复制序列 NovoBuilder中查看图谱

质粒操作方法

1. 发货形式：质粒干粉(常温运输，存于-20度，请务必先转化提质粒后使用)

2. 收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O溶解质粒;（质粒复测的浓度有时候与标称值差距较大，这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致，因此建议先转化提质粒后再使用）

3. 取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；

4. 加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min (30℃培养1-1.5小时);

5. 6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上;

6. 将平板倒置37℃培养14h，如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化；没有菌落则加入10μl质粒转化；建议不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态);

7. 挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。

pGWB702omega 载体序列

下载GenBank文件(.gb)

LOCUS       40924_23664       12156 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway binary vector pGWB702omega DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12156)
  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 12156)
  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 12156)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12156)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosci. 
            Biotechnol. Biochem."; date: "2011"; volume: "75"; issue: "4"; 
            pages: "804-807"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-JAN-2011) Contact:Tsuyoshi Nakagawa Shimane University, Center 
            for Integrated Research in Science; 1060 Nishikawatsu, Matsue, 
            Shimane 690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pPZP221.
FEATURES             Location/Qualifiers
     source          1..12156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(54..78)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        405..1083
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    1091..1157
                     /label=omega translational enhancer
                     /note="omega translational enhancer"
     protein_bind    1172..1296
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1333..1363
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1417..2073
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2418..2720
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2764..2888)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2920..3172
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3206..3222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3230..3246
                     /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(3254..3284)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3299..3320)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     terminator      complement(3356..3608)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(3646..4941)
                     /codon_start=1
                     /gene="gpt"
                     /product="UDP-N-acetylglucosamine: dolichol phosphate 
                     N-acetylglucosamine-1-P transferase"
                     /label=gpt
                     /protein_id="BAJ61164.1"
                     /translation="MAARKRASSISIPNKPDPSEPNSAPSEQKMTRKTVSASGEEFRLA
                     PPKLGVIFVISTLLCSLYLYLLCFHYKVDNELKRSILINAGLSLVGFFVTLKMIPVTAR
                     YVLRRNMFGFDINKRGTPQGDIKVPESLGIVVGIVFLIVAIIFQYFNFTEDSNWLVEYN
                     AALASICFMILLGFVDDVLDVPWRVKLVLPSFATLPLLMAYAGHTTIVIPKPLVAYIGL
                     EVLNLGRIYKLYMGLLAVFCTNSINIHAGLNGLEIGQTVVIAAAILIHNVMQIGASVDP
                     EYHQAHAFSIFLTQPLMATSLAMLAYNWYPSSVFVGDTYTVFAGMTMAVVGILGHFSET
                     LLIFFLPQVLNLLLSLPQLAGIVKCPRHRLPRYDPATGLLTGTKDGTLVNVYLRLFGPK
                     SEKSLCIHLLVFQALACAFCFILRHFLAGWYK"
     gene            complement(3646..4941)
                     /gene="gpt"
                     /label=gpt
     promoter        complement(4993..5172)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    complement(5678..5702)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6223..7011
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7260..7848
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(8034..8174)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8518..8712)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8781..9851)
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(10283..10909)
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"

pGWB702omega 载体 (V005691)

pGWB702omega 载体图谱

质粒操作方法

pGWB702omega 载体序列