■ 基本信息
启动子: | Tac |
复制子: | pBR322 |
终止子: | rrnB T1 |
质粒分类: | 大肠杆菌载体；蛋白表达质粒 |
质粒大小: | 4584bp |
原核抗性: | Amp |
克隆菌株: | DH5a |
培养条件: | 37度，37℃，有氧，LB |
表达宿主: | 大肠杆菌 |
诱导方式: | IPTG或乳糖及其类似物 |
备注: | 表达质粒 |
■ 质粒属性
质粒宿主: | 大肠杆菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 空载体 |
原核抗性: | Amp |
真核抗性: |
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荧光标记: |
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■ 质粒简介
pKK223-3 (1) contains the strong tac promoter upstream from the multiple cloning site and the strong rrnB ribosomal terminator downstream for control of protein expression (2-4). Induction: tac promoter is inducible with 1-5 mM IPTG. Promoter is very strong; even uninduced cells may show a low level of expression. Expression: Genes containing a ribosome-binding site and ATG can be inserted into any unique site in the MCS for expression. The ribosomebinding site on the plasmid can be utilised to express inserts if the ATG start codon of the insert is within 5-13 bp from the provided ribosomebindingsite (5, 6). Transcription terminators: rrnB transcription terminators stabilize the plasmid by inhibiting read-through transcription initiated from the tac promoter in the parent plasmid. Host(s): lac Iq strains; E. coli JM105 is recommended. Selectable marker(s): Plasmid confers resistance to 100 μg/ml ampicillin. Amplification: Recommended. Control Regions: Expression control region: tac promoter: -10: 50-44; -35: 72-67; Ribosome binding site: 11. MCS: 4552-4587. rrnB operon region: 5S rRNA region: 4471-4352; rrnB T1 terminator: 4348-4305; rrnB T2 terminator: 4173-4146. b-lactamase gene region: Promoter: -10: 4082-4077; -35: 4105-4100; Start codon (ATG): 4035; Stop codon (TAA): 3178. Plasmid replication region: Site of replication initiation: 2417-2415; Region necessary for replication: 3111-2415.
■ 质粒图谱
■ 质粒序列
质粒序列请下载：
ZK1616 pKK223-3大肠表达质粒.txt
