■ 產(chǎn)品簡(jiǎn)介
零背景ZT4-Blunt(Kan+)克隆試劑盒是專門對(duì)平滑末端DNA片段或PCR擴(kuò)增產(chǎn)物進(jìn)行高效克隆的一種陽(yáng)性篩選系統(tǒng)。該克隆載體包含一個(gè)致死基因，當(dāng)目的DNA片段插入克隆位點(diǎn)時(shí),該致死基因被破壞，故僅帶插入目的片段的克隆才能生長(zhǎng)形成白色菌落，無(wú)需加入IPTG及X-Gal進(jìn)行藍(lán)白斑篩選；克隆陽(yáng)性率超過95%。
■ 產(chǎn)品特點(diǎn)
·可5min快速連接，且克隆效率超過95%
·無(wú)需IPTG及X-Gal進(jìn)行藍(lán)白斑篩選
·PCR擴(kuò)增產(chǎn)物無(wú)需磷酸化
·酶切產(chǎn)物無(wú)需去磷酸化
·專用于平滑末端DNA片段或PCR擴(kuò)增產(chǎn)物
·特別適合Pfu、KOD等系列產(chǎn)物的克隆
■ 儲(chǔ)存
-20℃至少保存1年。
■ ZT4-Blunt(Kan+)載體測(cè)序引物序列
T7 Forward Sequencing Primer，20-mer：5’ -TAATACGACTCACTATAGGG-3’
RSP Reverse Sequencing Primer，24-mer：5’ -AAGAACATCGATTTTCCATGGCAG-3’
■ ZT4-Blunt(Kan+)載體圖譜
■ ZT4-Blunt(Kan+)載體多克隆位點(diǎn)序列
■ ZT4-Blunt(Kan+)載體全序列
1 GCCCCTGCAG CCGAATTATA TTATTTTTGC CAAATAATTT TTAACAAAAG
51 CTCTGAAGTC TTCTTCATTT AAATTCTTAG ATGATACTTC ATCTGGAAAA
101 TTGTCCCAAT TAGTAGCATC ACGCTGTGAG TAAGTTCTAA ACCATTTTTT
151 TATTGTTGTA TTATCTCTAA TCTTACTACT CGATGAGTTT TCGGTATTAT
201 CTCTATTTTT AACTTGGAGC AGGTTCCATT CATTGTTTTT TTCATCATAG
251 TGAATAAAAT CAACTGCTTT AACACTTGTG CCTGAACACC ATATCCATCC
301 GGCGTAATAC GACTCACTAT AGGGAGAGCG GCCGCCAGAT CTTCCGGATG
351 GCTCGAGTTT TTCAGCAAGA TATCTTTCTA GAAGATCTCC TACAATATTC
401 TCAGCTGCCA TGGAAAATCG ATGTTCTTCT TTTATTCTCT CAAGATTTTC
451 AGGCTGTATA TTAAAACTTA TATTAAGAAC TATGCTAACC ACCTCATCAG
501 GAACCGTTGT AGGTGGCGTG GGTTTTCTTG GCAATCGACT CTCATGAAAA
551 CTACGAGCTA AATATTCAAT ATGTTCCTCT TGACCAACTT TATTCTGCAT
601 TTTTTTTGAA CGAGGTTTAG AGCAAGCTTC AGGAAACTGA GACAGGAATT
651 TTATTAAAAA TTTAAATTTT GAAGAAAGTT CAGGGTTAAT AGCATCCATT
701 TTTTGCTTTG CAAGTTCCTC AGCATTCTTA ACAAAAGACG TCTCTTTTGA
751 CATGTTTAAA GTTTAAACCT CCTGTGTGAA ATTGTTATCC GCTCACAATT
801 CCACACATTA TACGAGCCGG AAGCATAAAG TGTAAAGCCT GGGGTGCCTA
851 ATGAGTGAGC TAACTCACAT TAATTGCGTT GCGCTCACTG CCAATTGCTT
901 TCCAGTCGGG AAACCTGTCG TGCCAGCTGC ATTAATGAAT CGGCCAACGC
951 GCGGGGAGAG GCGGTTTGCG TATTGGGCGC TCTTCCGCTT CCTCGCTCAC
1001 TGACTCGCTG CGCTCGGTCG TTCGGCTGCG GCGAGCGGTA TCAGCTCACT
1051 CAAAGGCGGT AATACGGTTA TCCACAGAAT CAGGGGATAA CGCAGGAAAG
1101 AACATGTGAG CAAAAGGCCA GCAAAAGGCC AGGAACCGTA AAAAGGCCGC
1151 GTTGCTGGCG TTTTTCCATA GGCTCCGCCC CCCTGACGAG CATCACAAAA
1201 ATCGACGCTC AAGTCAGAGG TGGCGAAACC CGACAGGACT ATAAAGATAC
1251 CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG TTCCGACCCT
1301 GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC
1351 TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC
1401 TCCAAGCTGG GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC
1451 CTTATCCGGT AACTATCGTC TTGAGTCCAA CCCGGTAAGA CACGACTTAT
1501 CGCCACTGGC AGCAGCCACT GGTAACAGGA TTAGCAGAGC GAGGTATGTA
1551 GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG GCTACACTAG
1601 AAGGACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA
1651 AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT
1701 GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA
1751 AGAAGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA
1801 ACTCACGTTA AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC
1851 TAGATCCTTT TAAATTAAAA ATGAAGTTTT AAATCAATCT AAAGTATATA
1901 TGAGTAAACT TGGTCTGACA GCTAAAACAA TTCATCCAGT AAAATATAAT
1951 ATTTTATTTT CTCCCAATCA GGCTTGATCC CCAGTAAGTC AAAAAATAGC
2001 TCGACATACT GTTCTTCCCC GATATCCTCC CTGATCGACC GGACGCAGAA
2051 GGCAATGTCA TACCACTTGT CCGCCCTGCC GCTTCTCCCA AGATCAATAA
2101 AGCCACTTAC TTTGCCATCT TTCACAAAGA TGTTGCTGTC TCCCAGGTCG
2151 CCGTGGGAAA AGACAAGTTC CTCTTCGGGC TTTTCCGTCT TTAAAAAATC
2201 ATACAGCTCG CGCGGATCTT TAAATGGAGT GTCTTCTTCC CAGTTTTCGC
2251 AATCCACATC GGCCAGATCG TTATTCAGTA AGTAATCCAA TTCGGCTAAG
2301 CGGCTGTCTA AGCTATTCGT ATAGGGACAA TCCGATATGT CGATGGAGTG
2351 AAAGAGCCTG ATGCACTCCG CATACAGCTC GATAATCTTT TCAGGGCTTT
2401 GTTCATCTTC ATACTCTTCC GAGCAAAGGA CGCCATCGGC CTCACTCATG
2451 AGCAGATTGC TCCAGCCATC ATGCCGTTCA AAGTGCAGGA CCTTTGGAAC
2501 AGGCAGCTTT CCTTCCAGCC ATAGCATCAT GTCCTTTTCC CGTTCCACAT
2551 CATAGGTGGT CCCTTTATAC CGGCTGTCCG TCATTTTTAA ATATAGGTTT
2601 TCATTTTCTC CCACCAGCTT ATATACCTTA GCAGGAGACA TTCCTTCCGT
2651 ATCTTTTACG CAGCGGTATT TTTCGATCAG TTTTTTCAAT TCCGGTGATA
2701 TTCTCATTTT AGCCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT
2751 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA
2801 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG
2851 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC
2901 ACGAGGCC
