SBB10Ntbk-DanielSedor
From OpenWetWare
Construction Files
sbb10
Construction of sbb10 PCR DS001/CA1600 on Bca1587 5-1 (420bp, gp=A) PCR CA1597/DS002 on Bca1587 5-6 (767bp, gp=B) ---------------------------- PCR DS001 and DS002 on A+B (1051 bp, EcoRI/BamHI) Sub into pBjk2741 (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-sb10 {<SB100x!} ---------------------------- DS001 Forward oligo for cloning of <SB100x! CCATAgaattcATGagatctGGTAAATCTAAAGAAATCTCTCAGGAC CA1600 CGAAACGCAGACGAGCTTTTTTGTGACGGTTCTGGAGCAGCGGTTTTTT PCA assembly of sleepingBeauty (Bca1587) CA1597 ATGCTGGAAGAAACCGGTACCAAAGTTTCTATCTCTACCGTTAAACGTG PCA assembly of sleepingBeauty (Bca1587) DS002 Reverse oligo for cloning of <SB100x! ctgatGGATCCttaGTATTTGGTAGCGTTACCTT Part: gatctGGTAAATCTAAAGAAATCTCTCAGGACCTGCGTAAACGTATCGTTGACCTGCACAAATCTGGTTCTTCTCTGGGTGCTATCTCTAAACGTCTGGCTGTTCCGCGTTCTTCTGTTCAGACCATCGTTCGTAAATACAAACA CCACGGTACCACCCAGCCGTCTTACCGTTCTGGTCGTCGTCGTGTTCTGTCTCCGCGTGACGAACGTACCCTGGTTCGTAAAGTTCAGATCAACCCGCGTACCACCGCTAAAGACCTGGTTAAAATGCTGGAAGAAACCGGTACC AAAGTTTCTATCTCTACCGTTAAACGTGTTCTGTACCGTCACAACCTGAAAGGTCACTCTGCTCGTAAAAAACCGCTGCTCCAGAACCGTCACAAAAAAGCTCGTCTGCGTTTCGCTACCGCTCACGGTGACAAAGACCGTACCT TCTGGCGTAACGTTCTGTGGTCTGACGAAACCAAAATCGAACTGTTCGGTCACAACGACCACCGTTACGTTTGGCGTAAAAAAGGTGAAGCTTGCAAACCGAAAAACACCATCCCGACCGTTAAACACGGTGGTGGTTCTATCAT GCTGTGGGGTTGCTTCGCTGCTGGTGGTACCGGTGCTCTGCACAAAATCGACGGTATCATGGACGCTGTTCAGTACGTTGACATCCTGAAACAGCACCTGAAAACCTCTGTTCGTAAACTGAAACTGGGTCGTAAATGGGTTTTC CAGCACGACAACGACCCGAAACACACCTCTAAAGTTGTTGCTAAATGGCTGAAAGACAACAAAGTTAAAGTTCTGGAATGGCCGTCTCAGTCTCCGGACCTGAACCCGATCGAAAACCTGTGGGCTGAACTGAAAAAACGTGTTC GTGCTCGTCGTCCGACCAACCTGACCCAGCTGCACCAGCTGTGCCAGGAAGAATGGGCTAAAATCCACCCGACCTACTGCGAAAAACTGGTTGAAGGTTACCCGAAACGTCTGACCCAGGTTAAACAGTTCAAAGGTAACGCTAC CAAATACTAAG
Relevant Protocols
SOEing PCR
Cloning by PCR
Gene Synthesis from Oligos (PCA method)
Zymo Gel Purification
Ligation of EcoRI/BamHI digests
Transformation by heat-shock
Picking of Colonies
sbb39
Eco/Bam transfer pBjh1601CK-Bjh1380 Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-Bjh1380 {PmgtCB}
Relevant Protocols
EcoRI/BamHI Transfer
Miniprep purification of DNA
sbb40
Eco/Bam transfer pBjh1601CK-Bjh1858 Sub into pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-Bjh1858 {<NIS!}
Relevant Protocols
EcoRI/BamHI Transfer
Miniprep purification of DNA