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:
- pSFS2A
- 载体抗性:
- Chloramphenicol
- 载体长度:
- 7510 bp
- 拷贝数:
- High copy number
- 感受态:
- DH5alpha
- 培养温度:
- 37℃
pSFS2A 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。
pSFS2A 载体序列
LOCUS Exported 7510 bp DNA circular SYN 23-DEC-2024
DEFINITION Cloning vector pSFS2A-mNeonGreen, complete sequence.
ACCESSION MK431400
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7510)
AUTHORS Mancera E, Frazer C, Porman AM, Ruiz-Castro S, Johnson AD, Bennett
RJ.
TITLE Genetic Modification of Closely Related Candida Species
JOURNAL Front Microbiol 10, 357 (2019)
PUBMED 30941104
REFERENCE 2 (bases 1 to 7510)
AUTHORS Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
TITLE Direct Submission
JOURNAL Submitted (23-JAN-2019) Departmento de Ingenieria Genetica, Centro
de Investigacion y de Estudios Avanzados del Instituto Politecnico
Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera
Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico
REFERENCE 3 (bases 1 to 7510)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7510)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Front
Microbiol 10, 357 (2019)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(23-JAN-2019) Departmento de Ingenieria Genetica, Centro de
Investigacion y de Estudios Avanzados del Instituto Politecnico
Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera
Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7510
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(915..7510,1..914)
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(142..246)
/label=AmpR promoter
rep_origin 273..729
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 870..886
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 896..914
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind complement(944..977)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
misc_feature 982..1529
/label=promoter region from Candida albicans MAL2
/note="promoter region from Candida albicans MAL2"
CDS 1536..2804
/label=FLP
/note="site-specific recombinase"
misc_feature 2808..3195
/label=terminator region from Candida albicans ACT1
/note="terminator region from Candida albicans ACT1"
misc_feature 3202..3699
/label=promoter region from Candida albicans ACT1
/note="promoter region from Candida albicans ACT1"
gene 3700..4926
/gene="SAT1"
/label=SAT1
CDS join(3700..3709,4364..4926)
/codon_start=1
/transl_table=11
/gene="SAT1"
/product="streptomycin acetyltransferase"
/label=SAT1
/note="nourseothricin resistance marker"
/protein_id="QBY25799.1"
/translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
FTYKTRPQVSNETAMYWYWFSGAQDDA"
misc_feature 4930..5059
/label=terminator region from Candida albicans URA3
/note="terminator region from Candida albicans URA3"
protein_bind complement(5064..5097)
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
primer_bind complement(5099..5115)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(5152..5170)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5191..5207)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5215..5231)
/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(5239..5269)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5284..5305)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5593..6181)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 6401..6503
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 6504..7160
/label=CmR
/note="chloramphenicol acetyltransferase"