测序及更多信息：https://pan.baidu.com/s/1YnZMKVRPFd6UoTzSPu-sWQ 密码：d2q1
采购地址：淘宝店铺地址
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载体名称:
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pKD46
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质粒类型:
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细菌表达；基因敲除系统
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克隆方法:
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多克隆位点，限制性内切酶
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载体大小:
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6329
bp
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备注:
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pKD46
is more efficient than pKD20 for making gene disruptions.复制子 pSC101 ori只能在30℃才能正常生长。
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Red同源重组系统由λ噬菌体exo、bet、gam三个基因组成，它们分别编码Exo、Beta、Gam三种蛋白质。
1、Exo蛋白
Exo蛋白是一种核酸外切酶，单亚基的分子量为24 kD，Exo蛋白的活性形式是一种环状三聚物分子，中间有一中空的通道，通道的一端可容纳双链DNA分子，另一端只可容纳单链DNA。Exo蛋白可结合在双链DNA的末端，从DNA双链的5’端向3’端降解DNA，使DNA分子形成3’粘性末端。
2、Beta蛋白
Beta蛋白在Red同源重组中起着决定性作用，具有介导互补单链DNA退火的功能，是一种退火蛋白，单亚基的分子量为25.8 kD。在溶液中，Beta蛋白自发地形成环状结构，紧紧地结合在单链DNA的3’突出端，防止DNA被单链核酸酶降解，并介导互补单链DNA的退火。双链DNA退火完成后，Beta蛋白从DNA双链上解离下来。
3、Gam蛋白
Gam蛋白为16kD的多肽分子，可与RecBCD核酸外切酶结合，抑制其对外源DNA的降解作用，防止外源DNA 进入细胞后被宿主降解。
全序列
1 CATCGATTTA TTATGACAAC TTGACGGCTA CATCATTCAC TTTTTCTTCA CAACCGGCAC
61 GGAACTCGCT CGGGCTGGCC CCGGTGCATT TTTTAAATAC CCGCGAGAAA TAGAGTTGAT
121 CGTCAAAACC AACATTGCGA CCGACGGTGG CGATAGGCAT CCGGGTGGTG CTCAAAAGCA
181 GCTTCGCCTG GCTGATACGT TGGTCCTCGC GCCAGCTTAA GACGCTAATC CCTAACTGCT
241 GGCGGAAAAG ATGTGACAGA CGCGACGGCG ACAAGCAAAC ATGCTGTGCG ACGCTGGCGA
301 TATCAAAATT GCTGTCTGCC AGGTGATCGC TGATGTACTG ACAAGCCTCG CGTACCCGAT
361 TATCCATCGG TGGATGGAGC GACTCGTTAA TCGCTTCCAT GCGCCGCAGT AACAATTGCT
421 CAAGCAGATT TATCGCCAGC AGCTCCGAAT AGCGCCCTTC CCCTTGCCCG GCGTTAATGA
481 TTTGCCCAAA CAGGTCGCTG AAATGCGGCT GGTGCGCTTC ATCCGGGCGA AAGAACCCCG
541 TATTGGCAAA TATTGACGGC CAGTTAAGCC ATTCATGCCA GTAGGCGCGC GGACGAAAGT
601 AAACCCACTG GTGATACCAT TCGCGAGCCT CCGGATGACG ACCGTAGTGA TGAATCTCTC
661 CTGGCGGGAA CAGCAAAATA TCACCCGGTC GGCAAACAAA TTCTCGTCCC TGATTTTTCA
721 CCACCCCCTG ACCGCGAATG GTGAGATTGA GAATATAACC TTTCATTCCC AGCGGTCGGT
781 CGATAAAAAA ATCGAGATAA CCGTTGGCCT CAATCGGCGT TAAACCCGCC ACCAGATGGG
841 CATTAAACGA GTATCCCGGC AGCAGGGGAT CATTTTGCGC TTCAGCCATA CTTTTCATAC
901 TCCCGCCATT CAGAGAAGAA ACCAATTGTC CATATTGCAT CAGACATTGC CGTCACTGCG
961 TCTTTTACTG GCTCTTCTCG CTAACCAAAC CGGTAACCCC GCTTATTAAA AGCATTCTGT
1021 AACAAAGCGG GACCAAAGCC ATGACAAAAA CGCGTAACAA AAGTGTCTAT AATCACGGCA
1081 GAAAAGTCCA CATTGATTAT TTGCACGGCG TCACACTTTG CTATGCCATA GCATTTTTAT
1141 CCATAAGATT AGCGGATCCT ACCTGACGCT TTTTATCGCA ACTCTCTACT GTTTCTCCAT
1201 ACCCGTTTTT TTGGGAATTC GAGCTCTAAG GAGGTTATAA AAAATGGATA TTAATACTGA
1261 AACTGAGATC AAGCAAAAGC ATTCACTAAC CCCCTTTCCT GTTTTCCTAA TCAGCCCGGC
1321 ATTTCGCGGG CGATATTTTC ACAGCTATTT CAGGAGTTCA GCCATGAACG CTTATTACAT
1381 TCAGGATCGT CTTGAGGCTC AGAGCTGGGC GCGTCACTAC CAGCAGCTCG CCCGTGAAGA
1441 GAAAGAGGCA GAACTGGCAG ACGACATGGA AAAAGGCCTG CCCCAGCACC TGTTTGAATC
1501 GCTATGCATC GATCATTTGC AACGCCACGG GGCCAGCAAA AAATCCATTA CCCGTGCGTT
1561 TGATGACGAT GTTGAGTTTC AGGAGCGCAT GGCAGAACAC ATCCGGTACA TGGTTGAAAC
1621 CATTGCTCAC CACCAGGTTG ATATTGATTC AGAGGTATAA AACGAATGAG TACTGCACTC
1681 GCAACGCTGG CTGGGAAGCT GGCTGAACGT GTCGGCATGG ATTCTGTCGA CCCACAGGAA
1741 CTGATCACCA CTCTTCGCCA GACGGCATTT AAAGGTGATG CCAGCGATGC GCAGTTCATC
1801 GCATTACTGA TCGTTGCCAA CCAGTACGGC CTTAATCCGT GGACGAAAGA AATTTACGCC
1861 TTTCCTGATA AGCAGAATGG CATCGTTCCG GTGGTGGGCG TTGATGGCTG GTCCCGCATC
1921 ATCAATGAAA ACCAGCAGTT TGATGGCATG GACTTTGAGC AGGACAATGA ATCCTGTACA
1981 TGCCGGATTT ACCGCAAGGA CCGTAATCAT CCGATCTGCG TTACCGAATG GATGGATGAA
2041 TGCCGCCGCG AACCATTCAA AACTCGCGAA GGCAGAGAAA TCACGGGGCC GTGGCAGTCG
2101 CATCCCAAAC GGATGTTACG TCATAAAGCC ATGATTCAGT GTGCCCGTCT GGCCTTCGGA
2161 TTTGCTGGTA TCTATGACAA GGATGAAGCC GAGCGCATTG TCGAAAATAC TGCATACACT
2221 GCAGAACGTC AGCCGGAACG CGACATCACT CCGGTTAACG ATGAAACCAT GCAGGAGATT
2281 AACACTCTGC TGATCGCCCT GGATAAAACA TGGGATGACG ACTTATTGCC GCTCTGTTCC
2341 CAGATATTTC GCCGCGACAT TCGTGCATCG TCAGAACTGA CACAGGCCGA AGCAGTAAAA
2401 GCTCTTGGAT TCCTGAAACA GAAAGCCGCA GAGCAGAAGG TGGCAGCATG ACACCGGACA
2461 TTATCCTGCA GCGTACCGGG ATCGATGTGA GAGCTGTCGA ACAGGGGGAT GATGCGTGGC
2521 ACAAATTACG GCTCGGCGTC ATCACCGCTT CAGAAGTTCA CAACGTGATA GCAAAACCCC
2581 GCTCCGGAAA GAAGTGGCCT GACATGAAAA TGTCCTACTT CCACACCCTG CTTGCTGAGG
2641 TTTGCACCGG TGTGGCTCCG GAAGTTAACG CTAAAGCACT GGCCTGGGGA AAACAGTACG
2701 AGAACGACGC CAGAACCCTG TTTGAATTCA CTTCCGGCGT GAATGTTACT GAATCCCCGA
2761 TCATCTATCG CGACGAAAGT ATGCGTACCG CCTGCTCTCC CGATGGTTTA TGCAGTGACG
2821 GCAACGGCCT TGAACTGAAA TGCCCGTTTA CCTCCCGGGA TTTCATGAAG TTCCGGCTCG
2881 GTGGTTTCGA GGCCATAAAG TCAGCTTACA TGGCCCAGGT GCAGTACAGC ATGTGGGTGA
2941 CGCGAAAAAA TGCCTGGTAC TTTGCCAACT ATGACCCGCG TATGAAGCGT GAAGGCCTGC
3001 ATTATGTCGT GATTGAGCGG GATGAAAAGT ACATGGCGAG TTTTGACGAG ATCGTGCCGG
3061 AGTTCATCGA AAAAATGGAC GAGGCACTGG CTGAAATTGG TTTTGTATTT GGGGAGCAAT
3121 GGCGATGACG CATCCTCACG ATAATATCCG GGTAGGCGCA ATCACTTTCG TCTACTCCGT
3181 TACAAAGCGA GGCTGGGTAT TTCCCGGCCT TTCTGTTATC CGAAATCCAC TGAAAGCACA
3241 GCGGCTGGCT GAGGAGATAA ATAATAAACG AGGGGCTGTA TGCACAAAGC ATCTTCTGTT
3301 GAGTTAAGAA CGAGTATCGA GATGGCACAT AGCCTTGCTC AAATTGGAAT CAGGTTTGTG
3361 CCAATACCAG TAGAAACAGA CGAAGAATCC ATGGGTATGG ACAGTTTTCC CTTTGATATG
3421 TAACGGTGAA CAGTTGTTCT ACTTTTGTTT GTTAGTCTTG ATGCTTCACT GATAGATACA
3481 AGAGCCATAA GAACCTCAGA TCCTTCCGTA TTTAGCCAGT ATGTTCTCTA GTGTGGTTCG
3541 TTGTTTTTGC GTGAGCCATG AGAACGAACC ATTGAGATCA TACTTACTTT GCATGTCACT
3601 CAAAAATTTT GCCTCAAAAC TGGTGAGCTG AATTTTTGCA GTTAAAGCAT CGTGTAGTGT
3661 TTTTCTTAGT CCGTTACGTA GGTAGGAATC TGATGTAATG GTTGTTGGTA TTTTGTCACC
3721 ATTCATTTTT ATCTGGTTGT TCTCAAGTTC GGTTACGAGA TCCATTTGTC TATCTAGTTC
3781 AACTTGGAAA ATCAACGTAT CAGTCGGGCG GCCTCGCTTA TCAACCACCA ATTTCATATT
3841 GCTGTAAGTG TTTAAATCTT TACTTATTGG TTTCAAAACC CATTGGTTAA GCCTTTTAAA
3901 CTCATGGTAG TTATTTTCAA GCATTAACAT GAACTTAAAT TCATCAAGGC TAATCTCTAT
3961 ATTTGCCTTG TGAGTTTTCT TTTGTGTTAG TTCTTTTAAT AACCACTCAT AAATCCTCAT
4021 AGAGTATTTG TTTTCAAAAG ACTTAACATG TTCCAGATTA TATTTTATGA ATTTTTTTAA
4081 CTGGAAAAGA TAAGGCAATA TCTCTTCACT AAAAACTAAT TCTAATTTTT CGCTTGAGAA
4141 CTTGGCATAG TTTGTCCACT GGAAAATCTC AAAGCCTTTA ACCAAAGGAT TCCTGATTTC
4201 CACAGTTCTC GTCATCAGCT CTCTGGTTGC TTTAGCTAAT ACACCATAAG CATTTTCCCT
4261 ACTGATGTTC ATCATCTGAG CGTATTGGTT ATAAGTGAAC GATACCGTCC GTTCTTTCCT
4321 TGTAGGGTTT TCAATCGTGG GGTTGAGTAG TGCCACACAG CATAAAATTA GCTTGGTTTC
4381 ATGCTCCGTT AAGTCATAGC GACTAATCGC TAGTTCATTT GCTTTGAAAA CAACTAATTC
4441 AGACATACAT CTCAATTGGT CTAGGTGATT TTAATCACTA TACCAATTGA GATGGGCTAG
4501 TCAATGATAA TTACTAGTCC TTTTCCTTTG AGTTGTGGGT ATCTGTAAAT TCTGCTAGAC
4561 CTTTGCTGGA AAACTTGTAA ATTCTGCTAG ACCCTCTGTA AATTCCGCTA GACCTTTGTG
4621 TGTTTTTTTT GTTTATATTC AAGTGGTTAT AATTTATAGA ATAAAGAAAG AATAAAAAAA
4681 GATAAAAAGA ATAGATCCCA GCCCTGTGTA TAACTCACTA CTTTAGTCAG TTCCGCAGTA
4741 TTACAAAAGG ATGTCGCAAA CGCTGTTTGC TCCTCTACAA AACAGACCTT AAAACCCTAA
4801 AGGCTTAAGT AGCACCCTCG CAAGCTCGGT TGCGGCCGCA ATCGGGCAAA TCGCTGAATA
4861 TTCCTTTTGT CTCCGACCAT CAGGCACCTG AGTCGCTGTC TTTTTCGTGA CATTCAGTTC
4921 GCTGCGCTCA CGGCTCTGGC AGTGAATGGG GGTAAATGGC ACTACAGGCG CCTTTTATGG
4981 ATTCATGCAA GGAAACTACC CATAATACAA GAAAAGCCCG TCACGGGCTT CTCAGGGCGT
5041 TTTATGGCGG GTCTGCTATG TGGTGCTATC TGACTTTTTG CTGTTCAGCA GTTCCTGCCC
5101 TCTGATTTTC CAGTCTGACC ACTTCGGATT ATCCCGTGAC AGGTCATTCA GACTGGCTAA
5161 TGCACCCAGT AAGGCAGCGG TATCATCAAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC
5221 ACGTTAAGGG ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA
5281 TTAAAAATGA AGTTTTAAAT CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA
5341 CCAATGCTTA ATCAGTGAGG CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT
5401 TGCCTGACTC CCCGTCGTGT AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG
5461 TGCTGCAATG ATACCGCGAG ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA
5521 GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC
5581 TATTAATTGT TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT
5641 TGTTGCCATT GCTACAGGCA TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG
5701 CTCCGGTTCC CAACGATCAA GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT
5761 TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT
5821 GGTTATGGCA GCACTGCATA ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT
5881 GACTGGTGAG TACTCAACCA AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC
5941 TTGCCCGGCG TCAATACGGG ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT
6001 CATTGGAAAA CGTTCTTCGG GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG
6061 TTCGATGTAA CCCACTCGTG CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT
6121 TTCTGGGTGA GCAAAAACAG GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG
6181 GAAATGTTGA ATACTCATAC TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA
6241 TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC
6301 GCGCACATTT CCCCGAAAAG TGCCACCTG
