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:
- pBIND-GR
- 载体抗性:
- Ampicillin
- 载体长度:
- 7548 bp
- 载体类型:
- Luciferase Vectors
- 复制子:
- ori
- 载体来源:
- Promega
- 筛选标记:
- Neomycin/G418(Geneticin)
- 拷贝数:
- High copy number
- 启动子:
- CMV
pBIND-GR 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。
pBIND-GR 载体序列
LOCUS 40924_6522 7548 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector pBIND-GR, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7548) AUTHORS Kopish K, Pagiuo A, TenHarmsel A. TITLE GloResponse(TM) 9XGAL4UAS-luc2P HEK293 Cell Line Technical Bulletin, TB552 JOURNAL Unpublished REFERENCE 2 (bases 1 to 7548) AUTHORS Kopish K, Pagiuo A, TenHarmsel A. TITLE Direct Submission JOURNAL Submitted (02-JUN-2009) Scientific Communications, Promega Corporation, 5500 E. Cheryl Parkway, Madison, WI 53711, USA REFERENCE 3 (bases 1 to 7548) TITLE Direct Submission REFERENCE 4 (bases 1 to 7548) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-JUN-2009) Scientific Communications, Promega Corporation, 5500 E. Cheryl Parkway, Madison, WI 53711, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7548 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 139..517 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 518..721 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" intron 857..989 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" promoter 1033..1051 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1083..1523 /codon_start=1 /label=GAL4 DNA binding domain /note="DNA binding domain of the GAL4 transcriptional activator" /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVA" misc_feature 1521..1553 /label=IgA linker /note="IgA linker" CDS 1563..2399 /codon_start=1 /gene="GR-LBD" /product="GR LBD" /label=GR-LBD /note="glucocorticoid receptor ligand binding domain" /protein_id="ACT53148.1" /translation="MQQATTGVSQETSENPGNKTIVPATLPQLTPTLVSLLEVIEPEVL YAGYDSSVPDSTWRIMTTLNMLGGRQVIAAVKWAKAIPGFRNLHLDDQMTLLQYSWMFL MAFALGWRSYRQSSANLLCFAPDLIINEQRMTLPCMYDQCKHMLYVSSELHRLQVSYEE YLCMKTLLLLSSVPKDGLKSQELFDEIRMTYIKELGKAIVKREGNSSQNWQRFYQLTKL LDSMHEVVENLLNYCFQTFLDKTMSIEFPEMLAEIITNQIPKYSNGNIKKLLFHQK" gene 1563..2399 /gene="GR-LBD" /label=GR-LBD misc_feature 2408..2463 /label=MCS /note="pUC18/19 multiple cloning site" polyA_signal complement(2568..2689) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 2934..3291 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3342..4274 /codon_start=1 /label=hRluc /note="Renilla luciferase" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" CDS 4326..5114 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 5181..5229 /label=poly(A) signal /note="synthetic polyadenylation signal" promoter 5385..5489 /label=AmpR promoter CDS 5490..6347 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin complement(6508..7096) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 7212..7495 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers"