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:
- pLIC-C-TAP
- 载体抗性:
- Streptomycin
- 载体长度:
- 11429 bp
- 载体类型:
- Expression vector
- 复制子:
- ori
- 载体来源:
- Popescu SC, Popescu GV, Bachan S, Zhang Z, Seay M, Gerstein M, Snyder M, Dinesh-Kumar SP.
- 启动子:
- CaMV35S(enhanced)
pLIC-C-TAP 载体图谱
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。
pLIC-C-TAP 载体序列
LOCUS 40924_28297 11429 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pLIC-C-TAP, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11429) AUTHORS Popescu SC, Popescu GV, Bachan S, Zhang Z, Seay M, Gerstein M, Snyder M, Dinesh-Kumar SP. TITLE Differential binding of calmodulin-related proteins to their targets revealed through high-density Arabidopsis protein microarrays JOURNAL Proc. Natl. Acad. Sci. U.S.A. 104 (11), 4730-4735 (2007) PUBMED 17360592 REFERENCE 2 (bases 1 to 11429) AUTHORS Popescu SC, Dinesh-Kumar SP. TITLE Direct Submission JOURNAL Submitted (17-JUL-2007) MCDB, Yale University, 219 Prospect Streeet, New Haven, CT 06520-8103, USA REFERENCE 3 (bases 1 to 11429) TITLE Direct Submission REFERENCE 4 (bases 1 to 11429) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl. Acad. Sci. U.S.A."; date: "2007"; volume: "104"; issue: "11"; pages: "4730-4735" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (17-JUL-2007) MCDB, Yale University, 219 Prospect Streeet, New Haven, CT 06520-8103, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..11429 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 737..1363 /codon_start=1 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 1800..2864 /codon_start=1 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" rep_origin 2933..3127 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 3471..3611 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(3797..4385) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4632..5420) /codon_start=1 /label=SmR /note="aminoglycoside adenylyltransferase (Murphy, 1985)" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" misc_feature 5948..5972 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal complement(6070..6244) /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(6282..6812) /codon_start=1 /label=GmR /note="gentamycin acetyltransferase" /translation="MVRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(6872..7550) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" primer_bind 7779..7795 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 7894..8567 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" misc_feature 8589..8644 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" protein_bind 8643..8667 /label=attB1 /note="recombination site for the Gateway(R) BP reaction" promoter 8831..8933 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 8964..9266 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(9320..9344) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" CDS 9382..9411 /codon_start=1 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" /translation="EQKLISEEDL" CDS 9418..9447 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9454..9483 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9502..9531 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9538..9567 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9574..9603 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9622..9651 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9658..9687 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9694..9723 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 9790..9807 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 9808..9831 /codon_start=1 /label=HRV 3C site /note="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /translation="LEVLFQGP" CDS 9871..10044 /codon_start=1 /label=ProtA /note="IgG-binding unit of Staphylococcus aureus protein A" /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL AEAKKLNDAQAPK" CDS 10048..10218 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNDAQAPK" terminator 10291..10543 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(10563..10579) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 10587..10603 /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(10611..10641) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(10656..10677) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." misc_feature 10842..10866 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA"