pETBlue-2 sequence landmarks 3606-3625 lac operator T7 promoter lac operator 22-
ID: 170070 • Letter: P
Question
pETBlue-2 sequence landmarks 3606-3625 lac operator T7 promoter lac operator 22-42 T7 transcription start multiple cloning region 276-467 (Nico HPac His Tag coding sequence 437-454 HSV. Tag coding sequence 395-430 lacz start codon 491 lacz peptide ORF 57-49 541-569 E. coli promoter fl origin 096-155 669-2526 bla coding sequence 3206 pUC origin. BspLU11 l (3263) Nsp 3267 AlwNI(2854) Rca I(2543 Xmn I (2338) Bcg I' (2278 Bcg 12244 Sca I (2219) Nco EcoRV 265) 13-ORF Sma I (290 Site Ecl136 II 295 Sac (297) BamH EcoRI (307) (313) PinA (3599) promoter Xba l Bg ll 223 Nhe I(235 Asc I (320 Dsa IC276) BssH 320) Pst I (330) Sse8387 (330) acO Sa 335) Aat II 336 Hinc ll (337 E. col promoter Cla I (348) AV Il 597 Mlu I 353 Nsp V (359) Hind III (362) Eag l (369) Pvu II (379 3-ORF Bst1107 l (384) Site Pml I (389) pETBlue-2 Xho I (431) Pac I (464) (3653 bp) Bpu1 102 BspE 1078) Dra III(1329) Ssp 534 Ahd I(1739)Explanation / Answer
The pETBlue-2 vectors are designed to identify recombinants by traditional blue/white screening while also allowing T7lac promoter based expression of target genes. Screening is independent of expression because the T7lac expression promoter is in an opposed orientation relative to the E. coli promoter that mediates blue/white screening. pETBlue-2 defines the open reading frame and inserts must be cloned in-frame if expression is desired. The vector features an expanded multiple cloning site (MCS) and optional C-terminal HSV-Tag and His-Tag sequences. The sequence is numbered from the first base of the T7 promoter sequence. The f1 origin in pETBlue-2 is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand.
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