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:
- pSB3K5-I52002
- 载体抗性:
- Kanamycin
- 载体长度:
- 4026 bp
- 载体类型:
- BioBrick cloning vector
- 复制子:
- p15A ori
- 载体来源:
- Shetty RP, Endy D, Knight TF Jr.
pSB3K5-I52002 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。
pSB3K5-I52002 载体序列
LOCUS 40924_38748 4026 bp DNA circular SYN 18-DEC-2018
DEFINITION BioBrick cloning vector pSB3K5-I52002, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4026)
AUTHORS Shetty RP, Endy D, Knight TF Jr.
TITLE Engineering BioBrick vectors from BioBrick parts
JOURNAL J Biol Eng 2, 5 (2008)
PUBMED 18410688
REFERENCE 2 (bases 1 to 4026)
AUTHORS Shetty RP, Endy D, Knight TF Jr.
TITLE Direct Submission
JOURNAL Submitted (15-FEB-2008) Biological Engineering, MIT, 32 Vassar
Street Room 32-311, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 4026)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4026)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Eng
2, 5 (2008)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-FEB-2008) Biological Engineering, MIT, 32 Vassar Street Room
32-311, Cambridge, MA 02139, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4026
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..21
/label=BioBrick suffix
/note="universal suffix for all parts"
misc_feature 35..46
/note="translational stop sequence for BioBrick
vector-insert insulation, has stop codons in all 6 reading
frames (BBa_B0042)"
terminator 57..88
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
primer_bind complement(116..135)
/note="VR reverse sequencing primer annealing site
(BBa_G00102)"
regulatory 136..176
/note="derived from ribosomal RNA rrnC operon, reverse
orientation (BBa_B0062); similar to Escherichia coli strain
MG1655 accession U00096"
/regulatory_class="terminator"
stem_loop complement(149..168)
/label=derived from ribosomal RNA rrnC operon
/note="derived from ribosomal RNA rrnC operon"
rep_origin 599..1143
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
CDS 1894..2706
/label=KanR
/note="aminoglycoside phosphotransferase"
terminator 2720..2779
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
primer_bind 2792..2811
/note="VF2 forward sequencing primer annealing site
(BBa_G00100)"
terminator complement(2839..2868)
/label=T3Te terminator
/note="phage T3 early transcription terminator"
misc_feature 2890..2901
/note="translational stop sequence for BioBrick
vector-insert insulation, has stop codons in all 6 reading
frames (BBa_B0042)"
misc_feature 2915..2936
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
'ATG'"
regulatory 2977..2982
/gene="ccdB"
/regulatory_class="minus_35_signal"
regulatory 3000..3005
/gene="ccdB"
/regulatory_class="minus_10_signal"
CDS 3044..3346
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
rep_origin 3379..3967
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"