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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃
- 保质期:
6个月
- 英文名:
pBR322
- 库存:
大量
- 供应商:
钰博生物
- 规格:
1ug/5ug
载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pBR322 |
| 质粒类型: | 克隆载体 |
| 高拷贝/低拷贝: | 低拷贝 |
| 克隆方法: | 限制性内切酶,多克隆位点 |
| 启动子: | 无 |
| 载体大小: | 4362 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | 无 |
| 载体抗性: | 四环素 |
| 克隆菌株: | DH5α 等 |
| 备注: | pBR322可作为载体骨架,构建各类载体 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
载体质粒图谱和多克隆位点信息

载体简介
载体序列
LOCUS pBR322 4361 bp DNA SYN
DEFINITION pBR322
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..4361
/organism="pBR322"
/mol_type="other DNA"
gene 86..1276
/label="tet"
/gene="tet"
CDS 86..1276
/label="ORF frame 2"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH
YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG
RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH
APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV
FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE
KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG
QLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR
ATST*"
misc_feature complement(438..457)
/label="pBRrevBam_primer"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH
YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG
RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH
APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV
FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE
KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG
QLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR
ATST*"
CDS complement(623..1081)
/label="ORF frame 2"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
misc_feature 1915..2106
/label="ROP"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
misc_feature complement(2100..2122)
/label="pGEX_3_primer"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
gene complement(3293..4153)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
CDS complement(3293..4153)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(4195..4223)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
361 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
421 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
481 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
541 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
601 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
661 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
721 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
781 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
841 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
901 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
961 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
1021 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
1081 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc
1141 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
1201 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
1261 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
1321 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
1381 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
1441 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
1501 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
1561 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
1621 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
1681 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
1741 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
1801 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
1861 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
1921 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
1981 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
2041 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
2101 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
2161 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
2221 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
2281 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
2341 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
2401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
2461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
2521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
2581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
2641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
2701 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
2761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
2821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
2881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
2941 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
3001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
3061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
3121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
3181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
3241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
3301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
3361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
3421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
3481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
3541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
3601 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
3661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
3721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
3781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
3841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
3901 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
3961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
4021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
4081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
4141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
4201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
4261 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
4321 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga a
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文献和实验Back to Basics: pBR322 and Protein Expression Systems in E. coli
plasmid-borne elements and their interaction with the particular host strain will influence the overall expression system and final productivity (3 ) (see Chapters 1 –3 ). Plasmid vector pBR322 (4 ) is a well-established multipurpose cloning
1),尿嘧啶合成酶基因(URA3),亮氨酸合成酶基因(LEU2),组氨酸合成酶基因(HIS4)等。 3) 生化标记基因。这类标记基因的表达产物可催化某些易检测的生化反应,常用的基因有β-半乳苷酶基因(lacZ)、葡萄糖苷酸酶基因(GUS)、氯霉素乙酰转移酶基因(CAT)。 为了进一步说明载体的结构和各个结构的功能,下面举一个在基因工程中使用比较频繁的大肠杆菌载体—pBR322作为例子。 大肠杆菌载体pBR322 载体pBR322是由Bolivar 等人于1977年构建的。该载
都中止。但松弛型质粒仍继续进行复制。因此在增殖松弛型质粒时,总是要让宿主菌经氯霉素处理(在培养液中加入适量的氯霉素)抑制蛋白质的合成。图23-3 pBR322的基本结构(一)大肠杆菌质粒应用的最广泛的质粒载体是pBR322,它属松弛型质粒,有抗氨苄青霉素和抗四环素两个抗性基因可作为标记基因,有许多种常用的限制酶的切点。它全长4352个核苷酸,排列顺序已全部测定,基本结构如图23-3。当需要用pBR322来克隆BamHI酶切的一个DNA片段时,一般的做法是如图23-4所示。图23-4 pBR322克隆
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