296:
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
253:
20:
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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
198:
resistance which allows for simpler screening process with antibiotic selection, but it is a low copy number plasmid which does not give a high yield of plasmid. Another plasmid, RSF 2124, which is a derivative of ColE1, confers ampicillin resistance but is larger.
202:
Many other plasmids were artificially constructed to create one that would be ideal for cloning purpose, and pBR322 was found to be most versatile by many and was therefore the one most popularly used. It has two antibiotic resistance genes, as
238:
contain the pBR322 origin of replication, and fragments of pBR322 are very popular in the construction of intraspecies shuttle or binary vectors and vectors for targeted integration and excision of DNA from chromosome.
511:
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977). "Construction and characterization of new cloning vehicles. II. A multipurpose cloning system".
476:
Bolivar F, Rodriguez RL, Betlach MC, Boyer HW (1977). "Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9".
547:
143:
site and the count increases through the Tet gene. If we have to remove ampicillin for instance, we must use restriction endonuclease or molecular scissors against
576:
Balbás P, Soberón X, Merino E, Zurita M, Lomeli H, Valle F, Flores N, Bolivar F (1986). "Plasmid vector pBR322 and its special-purpose derivatives--a review".
363:
Balbás P, Soberón X, Merino E, Zurita M, Lomeli H, Valle F, Flores N, Bolivar F (1986). "Plasmid vector pBR322 and its special-purpose derivatives--a review".
611:
Yanisch-Perron C, Vieira J, Messing J (1985). "Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors".
226:
A large number of other plasmids based on pBR322 have since been constructed specifically designed for a wide variety of purposes. Examples include the
147:
and then pBR322 will become anti-resistant to ampicillin. The same process of
Insertional Inactivation can be applied to Tetracycline. The Amp gene is
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51:
711:
706:
663:
560:
136:
inside the Amp gene.The source of these antibiotic resistance genes are from pSC101 for
Tetracycline and RSF2124 for Ampicillin.
158:
of two different DNA fragments to create pBR322. P2 is in the same region as P1, but it is on the opposite strand and initiates
154:. Promoters P1 and P3 are for the beta-lactamase gene. P3 is the natural promoter, and P1 is artificially created by the
43:
159:
70:
55:
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59:
63:
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120:. Eleven of these forty sites lie within the Tet gene. There are two sites for restriction enzymes
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amplification of plasmid to produce a high yield of plasmid, however screening for immunity to
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A schematic representation of the pBR322 vector with restriction sites indicated in blue.
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Please expand the section to include this information. Further details may exist on the
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178:. Each of these plasmids may have its advantages and disadvantages. For example, the
47:
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plasmid and its derivatives have the advantage of higher copy number and allow for
95:
207:, and a number of convenient unique restriction sites that made it suitable as a
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resistance gene of pSC101, the ampicillin resistance gene of RSF 2124, and the
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211:. The plasmid was constructed with genetic material from 3 main sources – the
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Early cloning experiments may be conducted using natural plasmids such the
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384:
349:
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The circular sequence is numbered such that 0 is the middle of the unique
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497:
190:
E1 is not technically simple. The plasmid pSC101, a natural plasmid from
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Watson, N. (1988). "A new revision of the sequence of plasmid pBR322".
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121:
87:
40:
36:
31:
433:
Manen D, Caro L (February 1991). "The replication of plasmid pSC101".
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66:. The p stands for "plasmid," and BR for "Bolivar" and "Rodriguez."
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19:
18:
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402:"pBR322 Nucleotide Sequences, NCBI Sequence Viewer v2.0"
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in the direction of the tetracycline resistance gene.
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Paulina Balbás; Argelia
Lorence, eds. (April 2004).
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650:Recombinant Gene Expression: Reviews and Protocols
647:
546:S.B. Primrose & R.M Twyman (17 January 2006).
555:(7th ed.). Wiley-Blackwell. pp. 64–65.
69:pBR322 is 4361 base pairs in length and has two
8:
549:Principles of Gene Manipulation and Genomics
234:for extrachromosomal protein expression and
654:(2nd ed.). Humana Press Inc. pp.
98:resistance (Tet) protein. It contains the
264:about annotation: what basepairs are what.
310:List of restriction enzyme cutting sites
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52:University of California, San Francisco
46:. Created in 1977 in the laboratory of
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411:
396:
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34:and was one of the first widely used
7:
108:gene, which encodes a restrictor of
132:of the Tet gene. There are six key
447:10.1111/j.1365-2958.1991.tb02103.x
14:
219:of pMB1, a close relative of the
251:
422:(5th ed.). pp. 53–61.
420:Principles of Gene Manipulation
418:R.W. Old & S.B. Primrose.
1:
712:Molecular biology techniques
625:10.1016/0378-1119(85)90120-9
590:10.1016/0378-1119(86)90307-0
525:10.1016/0378-1119(77)90000-2
490:10.1016/0378-1119(77)90074-9
377:10.1016/0378-1119(86)90307-0
342:10.1016/0378-1119(88)90212-0
707:DNA mobile genetic elements
733:
287:The sequence in pBR322 is
230:series of plasmids. Most
112:. The plasmid has unique
56:Francisco Bolivar Zapata
60:postdoctoral researcher
262:is missing information
24:
100:origin of replication
71:antibiotic resistance
54:, it was named after
22:
404:. 30 September 2008.
217:replication elements
116:for more than forty
64:Raymond L. Rodriguez
118:restriction enzymes
110:plasmid copy number
232:expression vectors
205:selectable markers
25:
16:Artificial plasmid
301:
300:
285:
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192:Salmonella panama
134:restriction sites
114:restriction sites
102:of pMB1, and the
86:resistance (Amp)
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687:pBR322 Features
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90:, and the gene
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676:External links
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665:978-1592597741
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562:978-1405135443
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435:Mol. Microbiol
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209:cloning vector
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152:beta-lactamase
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619:(1): 103–19.
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584:(1–3): 3–40.
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260:This section
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160:transcription
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94:encoding the
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82:encoding the
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48:Herbert Boyer
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243:DNA sequence
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213:tetracycline
201:
196:tetracycline
191:
169:
138:
103:
96:tetracycline
91:
77:
73:genes – the
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35:
27:
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128:within the
701:Categories
692:pBR322 DNA
682:pBR322 Map
316:References
276:March 2019
194:, confers
166:Background
149:penicillin
84:ampicillin
268:talk page
223:plasmid.
717:Plasmids
463:37314534
304:See also
156:ligation
130:promoter
633:2985470
598:3034735
455:2041467
385:3034735
350:3063608
293:pBR322
188:colicin
122:HindIII
88:protein
50:at the
44:vectors
41:cloning
37:E. coli
32:plasmid
662:
631:
596:
559:
533:344137
531:
498:344136
496:
461:
453:
383:
348:
176:pSC101
58:, the
28:pBR322
658:–85.
553:(PDF)
459:S2CID
221:ColE1
180:ColE1
172:ColE1
141:EcoRI
30:is a
660:ISBN
629:PMID
613:Gene
594:PMID
578:Gene
557:ISBN
529:PMID
513:Gene
494:PMID
478:Gene
451:PMID
381:PMID
365:Gene
346:PMID
330:Gene
174:and
145:PstI
126:ClaI
124:and
92:tetA
75:gene
62:and
621:doi
586:doi
521:doi
486:doi
443:doi
373:doi
338:doi
228:pUC
105:rop
79:bla
703::
656:77
627:.
617:33
615:.
592:.
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527:.
515:.
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410:^
393:^
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Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
