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:
- pIBA105-strep-ERI1-deltaSAP
- 载体抗性:
- Ampicillin
- 载体长度:
- 6532 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Declercq M, Biquand E, Karim M, Pietrosemoli N
pIBA105-strep-ERI1-deltaSAP 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。
pIBA105-strep-ERI1-deltaSAP 载体序列
LOCUS 62056_12715 6532 bp DNA circular SYN 28-OCT-2020
DEFINITION Cloning vector pIBA105-strep-ERI1-deltaSAP, complete sequence.
ACCESSION MT966995
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6532)
AUTHORS Declercq M, Biquand E, Karim M, Pietrosemoli N, Jacob Y, Demeret C,
Barbezange C, van der Werf S.
TITLE Influenza A virus co-opts ERI1 exonuclease bound to histone mRNA to
promote viral transcription
JOURNAL Nucleic Acids Res 48 (18), 10428-10440 (2020)
PUBMED 32960265
REFERENCE 2 (bases 1 to 6532)
AUTHORS Declercq M, Biquand E, Demeret C, Karim M, Pietrosemoli N, Jacob Y,
Demeret C, Van der Werf S, Barbezange C.
TITLE Direct Submission
JOURNAL Submitted (03-SEP-2020) National Influenza Centre, Sciensano, Rue
Juliette Wytsman 14, Brussels 1180, Belgium
REFERENCE 3 (bases 1 to 6532)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res"; date: "2020"; volume: "48"; issue: "18"; pages:
"10428-10440"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-SEP-2020) National Influenza Centre, Sciensano, Rue Juliette
Wytsman 14, Brussels 1180, Belgium"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6532
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 832..852
/label=sequencing primer
/note="sequencing primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 904..927
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
CDS 964..987
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
misc_feature 988..993
/label=linker
/note="linker"
misc_feature 1063..2007
/note="exoribonuclease 1 with deleted SAP (SAF-box, Acinus
and PIAS) domain"
polyA_signal 2132..2356
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2402..2830
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2844..3173
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3240..4031
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 4208..4341
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4378..4394)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4402..4418)
/label=lac operator
/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(4426..4456)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4471..4492)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4780..5365)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5539..6396)
/label=AmpR
/note="beta-lactamase"
promoter complement(6397..6501)
/label=AmpR promoter