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:
- RS213_ErbB-RASER1N-OFP
- 载体抗性:
- Ampicillin
- 载体长度:
- 8785 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Chung HK, Zou X, Bajar BT, Brand VR
- 启动子:
- CMV
RS213_ErbB-RASER1N-OFP 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。
RS213_ErbB-RASER1N-OFP 载体序列
LOCUS V016574 8785 bp DNA circular SYN 18-MAY-2019
DEFINITION Exported.
ACCESSION V016574
VERSION V016574
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8785)
AUTHORS Chung HK, Zou X, Bajar BT, Brand VR, Huo Y, Alcudia JF, Ferrell JE
Jr., Lin MZ.
TITLE A compact synthetic pathway rewires cancer signaling to therapeutic
effector release
JOURNAL Science 364 (6439) (2019)
PUBMED 31048459
REFERENCE 2 (bases 1 to 8785)
AUTHORS Chung HK, Lin MZ.
TITLE Direct Submission
JOURNAL Submitted (16-APR-2019) Bioengineering, Stanford University, 269
Campus Drive, CCRS2100, Stanford, CA 94305, United States
REFERENCE 3 (bases 1 to 8785)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Science 364
(6439) (2019)"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-APR-2019) Bioengineering, Stanford University, 269 Campus Drive,
CCRS2100, Stanford, CA 94305, United States"
FEATURES Location/Qualifiers
source 1..8785
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 12..564
/label="nonstandard type: promoter; CMV_imm early promoter
2"
misc_feature 67..354
/label="CAG enhancer"
/note="CAG enhancer"
misc_feature 521..541
/label="nonstandard type: primer_bind; CMV fwd primer"
LTR 584..764
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 811..936
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1435..1668
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1853..1897
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 2046..2087
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2195..2312
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2371..2684
/label="U6 promoter"
/note="RNA polymerase III promoter for mouse U6 snRNA (Das
et al., 1988)"
primer_bind 2701..2717
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2759..2792)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
enhancer 2908..3211
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3212..3415
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3414..3465
/label="p1317"
/note="p1317"
misc_feature 3447..4004
/label="Shc-PTB"
/note="Shc-PTB"
misc_feature 3810..3830
/label="HIF1a"
/note="HIF1a"
CDS 4008..4031
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
misc_feature 4068..4646
/label="nonstandard type: Protease; NS3Tprotease"
misc_feature 4227..4229
/label="nonstandard type: misc; T54"
primer_bind 4394..4426
/label="p1278"
/note="p1278"
misc_feature complement(4412..4442)
/direction=LEFT
/label="nonstandard type: primer_bind_reverse; p1502 MreI
rev"
misc_feature 4647..4712
/label="P2A"
/note="P2A"
misc_feature complement(4653..4712)
/direction=LEFT
/label="nonstandard type: primer_bind_reverse; p1401 P2A
reverse"
CDS 4713..5363
/label="mKO2"
/note="monomeric Kusabira-Orange 2 fluorescent protein"
misc_feature 5376..5417
/label="nonstandard type: epitope; V5"
misc_feature 5418..5450
/label="nonstandard type: Substrate; substrate 4"
misc_feature complement(5451..5489)
/direction=LEFT
/label="nonstandard type: primer_bind_reverse; p1514 GGS
linker"
misc_feature 5469..5852
/label="nonstandard type: gBLOCK; HKC gBLOCK-5 VAV1"
misc_feature 5490..5822
/label="nonstandard type: SH2; VAV1"
primer_bind 5823..5858
/label="p1511 GGs linker before CAAX"
/note="p1511 GGs linker before CAAX"
misc_feature 5880..5939
/label="CAAX"
/note="CAAX"
misc_feature 5943..5948
/label="EcoRV"
/note="EcoRV"
misc_feature 5961..6549
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6552..6568)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 6734..6755
/label="U3PPT"
/note="U3PPT"
misc_feature 6734..6749
/label="cPPT(1)"
/note="cPPT(1)"
misc_feature 6791..6971
/label="truncHIV-1 3 LTR(1)"
/note="truncHIV-1 3 LTR(1)"
rep_origin complement(7033..7621)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7795..8652)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8653..8757)
/label="AmpR promoter"