SBB10Ntbk-ZhenZongHuang
From OpenWetWare
Zhen Z. Huang 15:05, 16 February 2010 (EST)
Planning for Wed, Feb 17th, 2010
sbb31
- PCR
- Analytical gel
- Zymol clean up
- digestion
- gel purification
- ligation
- mapping
- gel purification
sbb15
- Wobble
- Analytical gel
- Zymol small clean up
- digestion
- gel purification
- ligation
- mapping
- gel purification
Zhen Z. Huang 13:00, 18 February 2010 (EST)
Construction Files for my parts
sbb31
sbb31: CA42 origin of replication
Source: pEC22-CA42
Target Sequence: agcacttcagcgcgccgtagcatcgataaacattacgggatggggcgaaactgccatctgttcgaaatgacgcgcaaatgggcttacagggcgattcgtcagggctggccagcattctcacagtggcttgatgccgtgattcagcgtgtcgaaatgtacaacgcatcgcttcccgttccgctttcacctcctgaatgtcgggctattggcaagagtattgcgaaatacacgcacaggaacttcacggcggaaactttcgcacagtatgtggctgatacgcacacgccagaaatacaggccaagagaggcaggaaaggtggcatcgctaaaggcgaagcctacgatgacaagcgtttcatggcgctatgtatgctggagaatggatattctcagaaagctattgcggcgatgttggaggtttctactcgaaccattcgaaactggaaaagcggaaaatagcctatatcagataacagcgcctttctggcgtttttttgagcagtaggtcttttgccg
Vector: pBjk2741
Short description: oriCA42
Genbank reference: D30056.1
Family: Origin of Replication
PCR ZZH001 and ZZH002 on pEC22-CA42 (547 bp, EcoRI/BamHI)
Sub into pBca9523-Bca1144 (EcoRI/BamHI, 2472+910, L)
Product is pBca9523-sbb31 {oriCA42}
----------------------------
ZZH001 Forward oligo for cloning of oriCA42
ccataGAATTCatgAGATCTagcacttcagcgcgccgtag
ZZh002 Reverse oligo for cloning of oriCA42
ctgatGGATCCcggcaaaagacctactgctc
sbb15
sbb15: phiC31 attB
Source: Synthetic
Target Sequence: tgacggtctcgaagccgcggtgcgggtgccagggcgtgcccttgggctccccgggcgcgtactccacctcacccatctggtcca
Vector: pBjk2741
Short descriptions: phiC31 attB
Genbank reference: AB306970 (759…842)
Family: Phage att site
good 20 bp gtgccagggcgtgcccttgg
Wobble ZZH003/ZZH004 (115bp, EcoRI/BamHI)
Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 2170+910, L)
Product is pBjk2741-sbb15 {phiC31 attB}
----
ZZH003 Forward construction of phiC31 attB basic part
ccataGAATTCatgAGATCTtgacggtctcgaagccgcggtgcgggtgccagggcgtgcccttgg
ZZH004 Reverse construction of phiC31 attB basic part
ctgatGGATCCtggaccagatgggtgaggtggagtacgcgcccggggagcccaagggcacgccctggcac
