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pBluescript II SK(+) 质粒 (编号: V011757)

Note: pBluescript II SK(+) is one of the four multipurpose phagemid vectors, pBtuescriptll SK+, pBluescriptll SK-, pBluescriptll KS+, and pBluescriptll KS-, varying in the orientation of their polylinker (KS versus SK) and f1 origin (+ versus -). These vectors were designed to facilitate rapid mapping of DMA inserts.

pBluescript II SK(+)2961 bp600120018002400CAP binding sitelac promoterlac operatorM13 revT3 promoterMCST7 promoterM13 fwdf1 oriAmpR promoterAmpRori
复制序列 NovoBuilder中查看图谱

基本信息

载体名称:
pBluescript II SK(+)
载体抗性:
Ampicillin
载体长度:
2961 bp
载体类型:
I.M.A.G.E. Consortium Plasmids
复制子:
ori
载体来源:
Alting-Mees MA, Short JM.
拷贝数:
High copy number
5'引物:
M13 fwd
3'引物:
M13 rev
感受态:
DH10B
培养温度:
37℃

产品信息

质粒编号:V011757
质粒名称:pBluescript II SK(+)
规格:5 μg (冻干粉)

下载资源

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，供学术研究使用。

确保质粒的关键元件正确，但是我们并不能保证实验效果。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。

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。

参考文献

  • Ren W, Jiang Z, Zhang M, Kong L, Zhang H, Liu Y, Fu Q, Ma W. The chloroplast genome of Salix floderusii and characterization of chloroplast regulatory elements. Front Plant Sci. 2022 Aug 26;13:987443. doi: 10.3389/fpls.2022.987443. PMID: 36092427; PMCID: PMC9459086.

  • Aoki-Shioi N, Nagai Y, Deshimaru M, Terada S. Precursor genes of Bowman-Birk-type serine proteinase inhibitors comprise multiple inhibitory domains to promote diversity. Biochim Biophys Acta Gen Subj. 2023 Jan;1867(1):130248. doi: 10.1016/j.bbagen.2022.130248. Epub 2022 Oct 1. PMID: 36191739.

  • Wang J, Duan J, Zhu L, Jiang Z, Zhu G. Sequencing and generation of an infectious clone of the pathogenic goose parvovirus strain LH. Arch Virol. 2015 Mar;160(3):711-8. doi: 10.1007/s00705-014-2319-5. Epub 2015 Jan 7. PMID: 25559668.

pBluescript II SK(+) 质粒 (编号: V011757)序列

LOCUS       Exported                2961 bp DNA     circular SYN 30-SEP-2025
DEFINITION  Standard cloning vector (phagemid excised from lambda ZAPII). The f1
            (+) orientation allows rescue of sense strand ssDNA. pBluescript II 
            KS(+) has a reversed MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2961)
  AUTHORS   Alting-Mees MA, Short JM.
  TITLE     pBluescript II: gene mapping vectors.
  JOURNAL   Nucleic Acids Res. 1989;17:9494.
  PUBMED    2555794
REFERENCE   2  (bases 1 to 2961)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2961)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 2961)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 2961)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "1989"; volume: "17"; pages: "9494"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(633..2961,1..632)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1133..2961,1..1132)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    207..228
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        243..273
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    281..297
                     /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     305..321
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        342..360
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    373..480
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        489..507
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(517..533)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      675..1130
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1156..1260
                     /label=AmpR promoter
     CDS             1261..2118
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2292..2880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"