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:
- pCM168
- 载体抗性:
- Ampicillin
- 载体长度:
- 8074 bp
- 载体类型:
- Insertional cloning vector
- 复制子:
- ori
- 载体来源:
- Marx CJ, Lidstrom ME.
pCM168 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。
pCM168 载体序列
LOCUS 40924_11336 8074 bp DNA circular SYN 17-DEC-2018
DEFINITION Insertional cloning vector pCM168, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8074)
AUTHORS Marx CJ, Lidstrom ME.
TITLE Development of an insertional expression vector system for
Methylobacterium extorquens AM1 and generation of null mutants
lacking mtdA and/or fch
JOURNAL Microbiology (Reading, Engl.) 150 (PT 1), 9-19 (2004)
PUBMED 14702393
REFERENCE 2 (bases 1 to 8074)
AUTHORS Marx CJ, Lidstrom ME.
TITLE Direct Submission
JOURNAL Submitted (22-MAY-2003) Microbiology, University of Washington, Box
357242, Seattle, WA 98195, USA
REFERENCE 3 (bases 1 to 8074)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8074)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Microbiology (Reading, Engl.) 150 (PT 1), 9-19 (2004)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-MAY-2003) Microbiology, University of Washington, Box 357242,
Seattle, WA 98195, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8074
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 177..263
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 355..382
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 609..656
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS 883..1248
/codon_start=1
/gene="katA"
/allele="'katA"
/product="catalase"
/label=katA
/protein_id="AAP75625.1"
/translation="DGAMRTDGNHGAQVDYEPNSFGGPKQDPSFSEPPLRIQGDAGRYG
WPGDDEDLYGQPKLFWTKVLDEGGRKRLVENIVTSMGDSPRHIQERMIAHWFKVHEDFG
RGVAQGLGIDTAQAAAE"
gene 883..1248
/gene="katA"
/allele="'katA"
/label=katA
primer_bind complement(2049..2065)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2073..2089)
/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(2097..2127)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2142..2163)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2451..3039)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3213..4070)
/label=AmpR
/note="beta-lactamase"
promoter complement(4071..4175)
/label=AmpR promoter
CDS 4493..5390
/codon_start=1
/gene="katA"
/allele="katA'"
/product="catalase"
/label=katA
/protein_id="AAP75627.1"
/translation="MQDYHLIEKMAHFNRERIPERVVHAKGYGAYGTFRLTRSLSEFTR
AKVLTEVGKDVPMVARFSTVGGESGSADTARDPRGFALKFYTEEGNWDLVGNNTPIFFV
RDAIKFSDFIRTQKRDPRTHLKPHWRRWDFWSLSPEAIHQVMFLYSDRGTPKSARFMNG
YGSHTFSLWNDRGERHWVKFHFHTKQGIQNFSADEADEVAGKDPDYSSADLSTAIEQGD
FPKWKVSVQLMPELDADTYRINPFDLTKVWPHGDYPLIEIGEMELNRNPENYFAEIEQS
AFEPSNVVPGIGFSPHKM"
gene 4493..5390
/gene="katA"
/allele="katA'"
/label=katA
misc_recomb 5407..5440
/note="loxP; Cre-recombinase recognition site"
protein_bind 5407..5440
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(5796..6443)
/label=TetR
/note="tetracycline resistance regulatory protein"
CDS 6549..7745
/label=TcR
/note="tetracycline efflux protein"
protein_bind complement(7766..7799)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(7813..7829)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8053..8069
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"