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:
- pLenti6-HA-Blast
- 载体抗性:
- Ampicillin
- 载体长度:
- 8667 bp
- 载体类型:
- Lentiviral vector
- 复制子:
- ori
- 载体来源:
- De Groote P, Grootjans S, Lippens S, Eichperger C, Leurs K, Kahr I, Tanghe G, Bruggeman I, De Schamphelaire W, Urwyler C, Vandenabeele P, Haustraete J, Declercq W.
- 启动子:
- RSV
pLenti6-HA-Blast 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。
pLenti6-HA-Blast 载体序列
LOCUS V005058 8667 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V005058
VERSION V005058
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8667)
AUTHORS De Groote P, Grootjans S, Lippens S, Eichperger C, Leurs K, Kahr I,
Tanghe G, Bruggeman I, De Schamphelaire W, Urwyler C, Vandenabeele
P, Haustraete J, Declercq W.
TITLE Generation of a new Gateway-compatible inducible lentiviral vector
platform allowing easy derivation of co-transduced cells
JOURNAL BioTechniques 60 (5), 252-259 (2016)
PUBMED 27177818
REFERENCE 2 (bases 1 to 8667)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (22-DEC-2015) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 8667)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8667)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"BioTechniques"; date: "2016"; volume: "60"; issue: "5"; pages:
"252-259"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-DEC-2015) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8667
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..16
/label="HIV-1 cPPT"
/note="HIV-1 cPPT"
LTR 18..251
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 323..457
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 484..619
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(640..658)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(668..684)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 826..1281
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1307..1411
/label="AmpR promoter"
CDS 1412..2269
/label="AmpR"
/note="beta-lactamase"
rep_origin 2443..3031
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3319..3340
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3355..3385
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3393..3409
/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)."
primer_bind 3417..3433
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3454..3472
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3500..3726
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 3727..3907
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 3954..4079
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4572..4805
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 4990..5034
/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 5183..5224
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
enhancer 5313..5616
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 5617..5820
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 5937..6061
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 6086..6116
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
CDS 6170..6826
/label="CmR"
/note="chloramphenicol acetyltransferase"
CDS 7171..7473
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(7517..7641)
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 7685..7711
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
promoter 7756..8086
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 8134..8181
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 8200..8595
/label="BSD"
/note="blasticidin S deaminase"