■ 基本信息
啟動子: | Tac |
復制子: | pBR322 |
終止子: | rrnB T1 |
質粒分類: | 大腸桿菌載體；蛋白表達質粒 |
質粒大小: | 4584bp |
原核抗性: | Amp |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37度，37℃，有氧，LB |
表達宿主: | 大腸桿菌 |
誘導方式: | IPTG或乳糖及其類似物 |
備注: | 表達質粒 |
■ 質粒屬性
質粒宿主: | 大腸桿菌 |
質粒用途: | 蛋白表達 |
片段類型: | ORF |
片段物種: | 空載體 |
原核抗性: | Amp |
真核抗性: |
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熒光標記: |
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■ 質粒簡介
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.
■ 質粒圖譜
■ 質粒序列
質粒序列請下載：
ZK1616 pKK223-3大腸表達質粒.txt
