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质粒宝 pBeloBac11细菌人工染色体质粒 HH-Y-111
发布时间:2018-07-31 12:13:08 | 浏览次数:



测序及更多信息:https://pan.baidu.com/s/1LhkTexaSEV2MCjrkSjUkdw 
提取码:hb1w 

采购地址:淘宝店铺地址

pBeloBac11细菌人工染色体质粒 HH-Y-111



pBeloBAC11 is an E. coli plasmid cloning vector designed for the construction of Bacterial Artificial Chromosomes (BACs). It is maintained in single copy, which allows the cloning and stable maintenance of very large DNA fragments (up to 300 kb; 1).

Based on the Ori2 (OriS) replicon of the F (fertility) factor of E. coli, the vector encodes the SopAB functions for active partitioning (2). These functions act at SopC to ensure that each daughter cell gets a copy of the plasmid. Initiation factor RepE (also known as RepA) mediates assembly of a replication complex at Ori2 (3-5).

The cloning region includes the following features: unique cloning sites BamHI, SphI and HindIII in a lacZα gene allowing for insert screening by α-complementation; T7 and SP6 phage promoters reading into the cloning sites for generation of RNA probes for blot procedures; several GC-rich restriction sites flanking the cloning segment for removal of the cloned insert; a chloramphenicol selectable marker; a lambda cos site for packaging into phage lambda particles if desired and also assuring a unique cleavage site for mapping; and a loxP site for specific cleavage by Cre recombinase in the presence of loxP oligonucleotide (6).

Enzymes with unique restriction sites are shown in bold type. Location of sites of all NEB restriction enzymes can be found on the NEB web site (choose Technical Reference > DNA Sequences and Maps). Restriction site coordinates refer to the position of the 5´-most base on the top strand in each recognition sequence.

Open reading frame (ORF) coordinates are in the form "translational start translational stop; numbers refer to positions on the top (clockwise) strand, regardless of the direction of transcription and include the start and stop codons.

Lambda cos site coordinates are the boundaries of the HincII fragment surrounding the annealed 12 base overhangs.

 

 pBeloBac11细菌人工染色体质粒 HH-Y-111

  随着基因组学时代的到来,克隆并利用生物的重要基因,研究这些基因的结构、功能和进化已成为遗传学领域的热点之一。生物体的许多重要性状都涉及到复杂的生理生化反应,受多基因或基因簇的控制,与成百上万碱基的DNA片段相关,此外,复杂基因组的物理作图和基因的图位克隆也涉及到大片段的研究,因此提高载体的容纳量一直是克隆载体的重要发展方向之一。
        目前比较理想的细菌人工染色体(Bacterial artificial chromosomeBAC)是指一种以F质粒(F-plasmid)为基础建构而成的细菌染色体克隆载体,命名为pBACs,其容量在50 - 300 kb之间,各种类型的pBACs在大肠杆菌都呈单拷贝。
        BAC是以大肠杆菌致育因子(F因子)为基础的合成载体。F因子也称为F质粒,是一种性质粒,它可将宿主当然体基因转移到另一宿主细胞。F因子在大肠杆菌中的复制受到严格控制而保持低拷贝,一般为每个细胞1-2个拷贝,其parB基因能够排斥细胞中的外源性F质粒,限制了细胞内质粒分子间的重排,降低了重组BAC中外源DNA的重排水平和嵌合度。此外,F因子具有携带1M bp插入片段的潜能,这就使构具有大容量克隆能力的BAC载体成为可能。
        BAC载体主要包括oriSrepE(控制F质粒复制)和parAparB(控制拷贝数)等元件。以BAC为基础克隆的载体成嵌合体的频率较低,转化效率高,而且以环状结构存在于细菌体内,易于分离纯化,已被科学界广泛接受。目前主要用于大片段基因组文库的构建和大的基因簇(gene cluster)的相关研究,在各类生物基因组计划中发挥重要作用。


全序列

1        GCGGCCGCAA GGGGTTCGCG TCAGCGGGTG TTGGCGGGTG TCGGGGCTGG CTTAACTATG
61       CGGCATCAGA GCAGATTGTA CTGAGAGTGC ACCATATGCG GTGTGAAATA CCGCACAGAT
121      GCGTAAGGAG AAAATACCGC ATCAGGCGCC ATTCGCCATT CAGGCTGCGC AACTGTTGGG
181      AAGGGCGATC GGTGCGGGCC TCTTCGCTAT TACGCCAGCT GGCGAAAGGG GGATGTGCTG
241      CAAGGCGATT AAGTTGGGTA ACGCCAGGGT TTTCCCAGTC ACGACGTTGT AAAACGACGG
301      CCAGTGAATT GTAATACGAC TCACTATAGG GCGAATTCGA GCTCGGTACC CGGGGATCCT
361      CTAGAGTCGA CCTGCAGGCA TGCAAGCTTG AGTATTCTAT AGTGTCACCT AAATAGCTTG
421      GCGTAATCAT GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC
481      AACATACGAG CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC
541      ACATTAATTG CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG
601      CATTAATGAA TCGGCCAACG CGAACCCCTT GCGGCCGCCC GGGCCGTCGA CCAATTCTCA
661      TGTTTGACAG CTTATCATCG AATTTCTGCC ATTCATCCGC TTATTATCAC TTATTCAGGC
721      GTAGCAACCA GGCGTTTAAG GGCACCAATA ACTGCCTTAA AAAAATTACG CCCCGCCCTG
781      CCACTCATCG CAGTACTGTT GTAATTCATT AAGCATTCTG CCGACATGGA AGCCATCACA
841      AACGGCATGA TGAACCTGAA TCGCCAGCGG CATCAGCACC TTGTCGCCTT GCGTATAATA
901      TTTGCCCATG GTGAAAACGG GGGCGAAGAA GTTGTCCATA TTGGCCACGT TTAAATCAAA
961      ACTGGTGAAA CTCACCCAGG GATTGGCTGA GACGAAAAAC ATATTCTCAA TAAACCCTTT
1021     AGGGAAATAG GCCAGGTTTT CACCGTAACA CGCCACATCT TGCGAATATA TGTGTAGAAA
1081     CTGCCGGAAA TCGTCGTGGT ATTCACTCCA GAGCGATGAA AACGTTTCAG TTTGCTCATG
1141     GAAAACGGTG TAACAAGGGT GAACACTATC CCATATCACC AGCTCACCGT CTTTCATTGC
1201     CATACGGAAT TCCGGATGAG CATTCATCAG GCGGGCAAGA ATGTGAATAA AGGCCGGATA
1261     AAACTTGTGC TTATTTTTCT TTACGGTCTT TAAAAAGGCC GTAATATCCA GCTGAACGGT
1321     CTGGTTATAG GTACATTGAG CAACTGACTG AAATGCCTCA AAATGTTCTT TACGATGCCA
1381     TTGGGATATA TCAACGGTGG TATATCCAGT GATTTTTTTC TCCATTTTAG CTTCCTTAGC
1441     TCCTGAAAAT CTCGATAACT CAAAAAATAC GCCCGGTAGT GATCTTATTT CATTATGGTG
1501     AAAGTTGGAA CCTCTTACGT GCCGATCAAC GTCTCATTTT CGCCAAAAGT TGGCCCAGGG
1561     CTTCCCGGTA TCAACAGGGA CACCAGGATT TATTTATTCT GCGAAGTGAT CTTCCGTCAC
1621     AGGTATTTAT TCGCGATAAG CTCATGGAGC GGCGTAACCG TCGCACAGGA AGGACAGAGA
1681     AAGCGCGGAT CTGGGAAGTG ACGGACAGAA CGGTCAGGAC CTGGATTGGG GAGGCGGTTG
1741     CCGCCGCTGC TGCTGACGGT GTGACGTTCT CTGTTCCGGT CACACCACAT ACGTTCCGCC
1801     ATTCCTATGC GATGCACATG CTGTATGCCG GTATACCGCT GAAAGTTCTG CAAAGCCTGA
1861     TGGGACATAA GTCCATCAGT TCAACGGAAG TCTACACGAA GGTTTTTGCG CTGGATGTGG
1921     CTGCCCGGCA CCGGGTGCAG TTTGCGATGC CGGAGTCTGA TGCGGTTGCG ATGCTGAAAC
1981     AATTATCCTG AGAATAAATG CCTTGGCCTT TATATGGAAA TGTGGAACTG AGTGGATATG
2041     CTGTTTTTGT CTGTTAAACA GAGAAGCTGG CTGTTATCCA CTGAGAAGCG AACGAAACAG
2101     TCGGGAAAAT CTCCCATTAT CGTAGAGATC CGCATTATTA ATCTCAGGAG CCTGTGTAGC
2161     GTTTATAGGA AGTAGTGTTC TGTCATGATG CCTGCAAGCG GTAACGAAAA CGATTTGAAT
2221     ATGCCTTCAG GAACAATAGA AATCTTCGTG CGGTGTTACG TTGAAGTGGA GCGGATTATG
2281     TCAGCAATGG ACAGAACAAC CTAATGAACA CAGAACCATG ATGTGGTCTG TCCTTTTACA
2341     GCCAGTAGTG CTCGCCGCAG TCGAGCGACA GGGCGAAGCC CTCGAGTGAG CGAGGAAGCA
2401     CCAGGGAACA GCACTTATAT ATTCTGCTTA CACACGATGC CTGAAAAAAC TTCCCTTGGG
2461     GTTATCCACT TATCCACGGG GATATTTTTA TAATTATTTT TTTTATAGTT TTTAGATCTT
2521     CTTTTTTAGA GCGCCTTGTA GGCCTTTATC CATGCTGGTT CTAGAGAAGG TGTTGTGACA
2581     AATTGCCCTT TCAGTGTGAC AAATCACCCT CAAATGACAG TCCTGTCTGT GACAAATTGC
2641     CCTTAACCCT GTGACAAATT GCCCTCAGAA GAAGCTGTTT TTTCACAAAG TTATCCCTGC
2701     TTATTGACTC TTTTTTATTT AGTGTGACAA TCTAAAAACT TGTCACACTT CACATGGATC
2761     TGTCATGGCG GAAACAGCGG TTATCAATCA CAAGAAACGT AAAAATAGCC CGCGAATCGT
2821     CCAGTCAAAC GACCTCACTG AGGCGGCATA TAGTCTCTCC CGGGATCAAA AACGTATGCT
2881     GTATCTGTTC GTTGACCAGA TCAGAAAATC TGATGGCACC CTACAGGAAC ATGACGGTAT
2941     CTGCGAGATC CATGTTGCTA AATATGCTGA AATATTCGGA TTGACCTCTG CGGAAGCCAG
3001     TAAGGATATA CGGCAGGCAT TGAAGAGTTT CGCGGGGAAG GAAGTGGTTT TTTATCGCCC
3061     TGAAGAGGAT GCCGGCGATG AAAAAGGCTA TGAATCTTTT CCTTGGTTTA TCAAACGTGC
3121     GCACAGTCCA TCCAGAGGGC TTTACAGTGT ACATATCAAC CCATATCTCA TTCCCTTCTT
3181     TATCGGGTTA CAGAACCGGT TTACGCAGTT TCGGCTTAGT GAAACAAAAG AAATCACCAA
3241     TCCGTATGCC ATGCGTTTAT ACGAATCCCT GTGTCAGTAT CGTAAGCCGG ATGGCTCAGG
3301     CATCGTCTCT CTGAAAATCG ACTGGATCAT AGAGCGTTAC CAGCTGCCTC AAAGTTACCA
3361     GCGTATGCCT GACTTCCGCC GCCGCTTCCT GCAGGTCTGT GTTAATGAGA TCAACAGCAG
3421     AACTCCAATG CGCCTCTCAT ACATTGAGAA AAAGAAAGGC CGCCAGACGA CTCATATCGT
3481     ATTTTCCTTC CGCGATATCA CTTCCATGAC GACAGGATAG TCTGAGGGTT ATCTGTCACA
3541     GATTTGAGGG TGGTTCGTCA CATTTGTTCT GACCTACTGA GGGTAATTTG TCACAGTTTT
3601     GCTGTTTCCT TCAGCCTGCA TGGATTTTCT CATACTTTTT GAACTGTAAT TTTTAAGGAA
3661     GCCAAATTTG AGGGCAGTTT GTCACAGTTG ATTTCCTTCT CTTTCCCTTC GTCATGTGAC
3721     CTGATATCGG GGGTTAGTTC GTCATCATTG ATGAGGGTTG ATTATCACAG TTTATTACTC
3781     TGAATTGGCT ATCCGCGTGT GTACCTCTAC CTGGAGTTTT TCCCACGGTG GATATTTCTT
3841     CTTGCGCTGA GCGTAAGAGC TATCTGACAG AACAGTTCTT CTTTGCTTCC TCGCCAGTTC
3901     GCTCGCTATG CTCGGTTACA CGGCTGCGGC GAGCGCTAGT GATAATAAGT GACTGAGGTA
3961     TGTGCTCTTC TTATCTCCTT TTGTAGTGTT GCTCTTATTT TAAACAACTT TGCGGTTTTT
4021     TGATGACTTT GCGATTTTGT TGTTGCTTTG CAGTAAATTG CAAGATTTAA TAAAAAAACG
4081     CAAAGCAATG ATTAAAGGAT GTTCAGAATG AAACTCATGG AAACACTTAA CCAGTGCATA
4141     AACGCTGGTC ATGAAATGAC GAAGGCTATC GCCATTGCAC AGTTTAATGA TGACAGCCCG
4201     GAAGCGAGGA AAATAACCCG GCGCTGGAGA ATAGGTGAAG CAGCGGATTT AGTTGGGGTT
4261     TCTTCTCAGG CTATCAGAGA TGCCGAGAAA GCAGGGCGAC TACCGCACCC GGATATGGAA
4321     ATTCGAGGAC GGGTTGAGCA ACGTGTTGGT TATACAATTG AACAAATTAA TCATATGCGT
4381     GATGTGTTTG GTACGCGATT GCGACGTGCT GAAGACGTAT TTCCACCGGT GATCGGGGTT
4441     GCTGCCCATA AAGGTGGCGT TTACAAAACC TCAGTTTCTG TTCATCTTGC TCAGGATCTG
4501     GCTCTGAAGG GGCTACGTGT TTTGCTCGTG GAAGGTAACG ACCCCCAGGG AACAGCCTCA
4561     ATGTATCACG GATGGGTACC AGATCTTCAT ATTCATGCAG AAGACACTCT CCTGCCTTTC
4621     TATCTTGGGG AAAAGGACGA TGTCACTTAT GCAATAAAGC CCACTTGCTG GCCGGGGCTT
4681     GACATTATTC CTTCCTGTCT GGCTCTGCAC CGTATTGAAA CTGAGTTAAT GGGCAAATTT
4741     GATGAAGGTA AACTGCCCAC CGATCCACAC CTGATGCTCC GACTGGCCAT TGAAACTGTT
4801     GCTCATGACT ATGATGTCAT AGTTATTGAC AGCGCGCCTA ACCTGGGTAT CGGCACGATT
4861     AATGTCGTAT GTGCTGCTGA TGTGCTGATT GTTCCCACGC CTGCTGAGTT GTTTGACTAC
4921     ACCTCCGCAC TGCAGTTTTT CGATATGCTT CGTGATCTGC TCAAGAACGT TGATCTTAAA
4981     GGGTTCGAGC CTGATGTACG TATTTTGCTT ACCAAATACA GCAATAGTAA TGGCTCTCAG
5041     TCCCCGTGGA TGGAGGAGCA AATTCGGGAT GCCTGGGGAA GCATGGTTCT AAAAAATGTT
5101     GTACGTGAAA CGGATGAAGT TGGTAAAGGT CAGATCCGGA TGAGAACTGT TTTTGAACAG
5161     GCCATTGATC AACGCTCTTC AACTGGTGCC TGGAGAAATG CTCTTTCTAT TTGGGAACCT
5221     GTCTGCAATG AAATTTTCGA TCGTCTGATT AAACCACGCT GGGAGATTAG ATAATGAAGC
5281     GTGCGCCTGT TATTCCAAAA CATACGCTCA ATACTCAACC GGTTGAAGAT ACTTCGTTAT
5341     CGACACCAGC TGCCCCGATG GTGGATTCGT TAATTGCGCG CGTAGGAGTA ATGGCTCGCG
5401     GTAATGCCAT TACTTTGCCT GTATGTGGTC GGGATGTGAA GTTTACTCTT GAAGTGCTCC
5461     GGGGTGATAG TGTTGAGAAG ACCTCTCGGG TATGGTCAGG TAATGAACGT GACCAGGAGC
5521     TGCTTACTGA GGACGCACTG GATGATCTCA TCCCTTCTTT TCTACTGACT GGTCAACAGA
5581     CACCGGCGTT CGGTCGAAGA GTATCTGGTG TCATAGAAAT TGCCGATGGG AGTCGCCGTC
5641     GTAAAGCTGC TGCACTTACC GAAAGTGATT ATCGTGTTCT GGTTGGCGAG CTGGATGATG
5701     AGCAGATGGC TGCATTATCC AGATTGGGTA ACGATTATCG CCCAACAAGT GCTTATGAAC
5761     GTGGTCAGCG TTATGCAAGC CGATTGCAGA ATGAATTTGC TGGAAATATT TCTGCGCTGG
5821     CTGATGCGGA AAATATTTCA CGTAAGATTA TTACCCGCTG TATCAACACC GCCAAATTGC
5881     CTAAATCAGT TGTTGCTCTT TTTTCTCACC CCGGTGAACT ATCTGCCCGG TCAGGTGATG
5941     CACTTCAAAA AGCCTTTACA GATAAAGAGG AATTACTTAA GCAGCAGGCA TCTAACCTTC
6001     ATGAGCAGAA AAAAGCTGGG GTGATATTTG AAGCTGAAGA AGTTATCACT CTTTTAACTT
6061     CTGTGCTTAA AACGTCATCT GCATCAAGAA CTAGTTTAAG CTCACGACAT CAGTTTGCTC
6121     CTGGAGCGAC AGTATTGTAT AAGGGCGATA AAATGGTGCT TAACCTGGAC AGGTCTCGTG
6181     TTCCAACTGA GTGTATAGAG AAAATTGAGG CCATTCTTAA GGAACTTGAA AAGCCAGCAC
6241     CCTGATGCGA CCACGTTTTA GTCTACGTTT ATCTGTCTTT ACTTAATGTC CTTTGTTACA
6301     GGCCAGAAAG CATAACTGGC CTGAATATTC TCTCTGGGCC CACTGTTCCA CTTGTATCGT
6361     CGGTCTGATA ATCAGACTGG GACCACGGTC CCACTCGTAT CGTCGGTCTG ATTATTAGTC
6421     TGGGACCACG GTCCCACTCG TATCGTCGGT CTGATTATTA GTCTGGGACC ACGGTCCCAC
6481     TCGTATCGTC GGTCTGATAA TCAGACTGGG ACCACGGTCC CACTCGTATC GTCGGTCTGA
6541     TTATTAGTCT GGGACCATGG TCCCACTCGT ATCGTCGGTC TGATTATTAG TCTGGGACCA
6601     CGGTCCCACT CGTATCGTCG GTCTGATTAT TAGTCTGGAA CCACGGTCCC ACTCGTATCG
6661     TCGGTCTGAT TATTAGTCTG GGACCACGGT CCCACTCGTA TCGTCGGTCT GATTATTAGT
6721     CTGGGACCAC GATCCCACTC GTGTTGTCGG TCTGATTATC GGTCTGGGAC CACGGTCCCA
6781     CTTGTATTGT CGATCAGACT ATCAGCGTGA GACTACGATT CCATCAATGC CTGTCAAGGG
6841     CAAGTATTGA CATGTCGTCG TAACCTGTAG AACGGAGTAA CCTCGGTGTG CGGTTGTATG
6901     CCTGCTGTGG ATTGCTGCTG TGTCCTGCTT ATCCACAACA TTTTGCGCAC GGTTATGTGG
6961     ACAAAATACC TGGTTACCCA GGCCGTGCCG GCACGTTAAC CGGGCTGCAT CCGATGCAAG
7021     TGTGTCGCTG TCGACGAGCT CGCGAGCTCG GACATGAGGT TGCCCCGTAT TCAGTGTCGC
7081     TGATTTGTAT TGTCTGAAGT TGTTTTTACG TTAAGTTGAT GCAGATCAAT TAATACGATA
7141     CCTGCGTCAT AATTGATTAT TTGACGTGGT TTGATGGCCT CCACGCACGT TGTGATATGT
7201     AGATGATAAT CATTATCACT TTACGGGTCC TTTCCGGTGA TCCGACAGGT TACGGGGCGG
7261     CGACCTCGCG GGTTTTCGCT ATTTATGAAA ATTTTCCGGT TTAAGGCGTT TCCGTTCTTC
7321     TTCGTCATAA CTTAATGTTT TTATTTAAAA TACCCTCTGA AAAGAAAGGA AACGACAGGT
7381     GCTGAAAGCG AGCTTTTTGG CCTCTGTCGT TTCCTTTCTC TGTTTTTGTC CGTGGAATGA
7441     ACAATGGAAG TCCGAGCTCA TCGCTAATAA CTTCGTATAG CATACATTAT ACGAAGTTAT
7501     ATTCGAT





 
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