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:
- pLB2 CAG P2Gm
- 载体抗性:
- Ampicillin
- 载体长度:
- 10656 bp
- 载体类型:
- Mammalian Expression, Mouse Targeting, Lentiviral,
- 复制子:
- ori
- 筛选标记:
- Puromycin
- 拷贝数:
- Low Copy
- 启动子:
- CAG
- 克隆方法:
- Restriction Enzyme
- 5'引物:
- GFP miR primer (GCTGGAGTTCGTGACCGCC)
pLB2 CAG P2Gm 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。
pLB2 CAG P2Gm 载体序列
LOCUS V000590 10656 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000590
VERSION V000590
KEYWORDS pLB2 CAG P2Gm
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10656)
AUTHORS Stern P, Astrof S, Erkeland SJ, Schustak J, Sharp PA, Hynes RO
TITLE A system for Cre-regulated RNA interference in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2008 Sep 16. 105(37):13895-900.
PUBMED 18779577
REFERENCE 2 (bases 1 to 10656)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 10656)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 2008 Sep 16. 105(37):13895-900."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10656
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(47..66)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 355..372
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 489..510
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 525..555
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 563..579
/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)."
primer_bind 568..590
/label="M13/pUC Reverse"
/note="In lacZ gene"
primer_bind 587..603
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 587..603
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
promoter 641..868
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 869..1049
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1096..1221
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1720..1953
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2138..2182
/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 2331..2372
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2480..2597
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 3119..3422
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3424..3701
/label="chicken beta-actin promoter"
intron 3703..4720
/label="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 4728..4747
/label="pCAG-F"
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 4809..4818
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4815..5411
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label="PuroR"
/note="confers resistance to puromycin"
/translation="MVEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 5311..5331
/label="Puro-F"
/note="Puromycin resistance gene, forward primer"
CDS 5433..5498
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label="F2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 5514..6230
/label="EGFP"
/note="enhanced GFP"
ncRNA 6283..6404
/label="5' miR-30a"
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
ncRNA 6431..6445
/label="miR-30a loop"
/note="loop from the the 71-nt precursor of the human
miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
ncRNA 6473..6599
/label="3' miR-30a"
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
misc_feature 6685..7273
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7276..7292)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7277..7293)
/label="pBluescriptKS"
/note="For pBluescript vector"
polyA_signal 8436..8643
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
LTR 8662..8842
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(8904..9492)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9666..10523)
/label="AmpR"
/note="beta-lactamase"
promoter complement(10524..10628)
/label="AmpR promoter"