pBR322
PVT0009 2ug
pBR322 Informaiton
Replicon: PBR322 ori
Plasmid classification: large intestine series plasmid; large intestine clone plasmid; gene clone plasmid.
Plasmid size: 4361bp
Prokaryotic resistance: ampicillin Amp (100 g/ml), tetracycline Tet (10 g/ml)
Cloning strains: E. coli DH5 and E.
Culture conditions: 37 C, aerobic, LB
5'sequencing primers: PBR322.F (AAGTGCCACCTGACGTCTAA)
3'sequencing primers: PBR322.R (GCCTGCCACCATACCCAC)
pBR322 Description
The pBR322 plasmid has a small relative molecular weight and has a replication initiation site, which ensures that the plasmid can only replicate in E. coli cells. There are two antibiotic resistance genes, ampicillin resistance gene and tetracycline resistance gene, which can be used as selective markers for transformants. With high copy number, each cell can accumulate 1000-3000 copies after chloramphenicol amplification, which provides great convenience for the preparation of recombinant DNA. At the same time, because of its artificial construction of plasmids, although with resistance genes, but can not transfer between natural host cells, and the use of safe strains, will not cause antibiotic resistance gene transmission.
PBR322 is an important plasmid constructed artificially and has universal plasmid. It was constructed by a complex recombination process of pSF2124, pMB1 and pSC101 three parental plasmids.
The size of plasmid vector pBR322 is 4361bp, and its relative molecular weight is the first advantage. The second advantage is that it carries a replication initiation site, which ensures that the plasmid can only replicate in E. coli cells. It has two antibiotic resistance genes, ampicillin resistance gene and tetracycline resistance gene. Selective markers for transformants are its third advantage.
pBR322 Multiple cloning site
pBR322 Sequence
LOCUS Exported 4361 bp ds-DNA circular SYN 22-9-2015
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4361)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-9-22
FEATURES Location/Qualifiers
source 1..4361
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..38
/note="tet promoter"
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 86..1276
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/note="TcR"
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
CDS 1915..2106
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2208..2348
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(2534..3122)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3293..4153)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4154..4258)
/gene="bla"
/note="AmpR promoter"
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
//
Caution
Product is for research use only!
Search name
pBR322,Plasmid pBR322,pBR322 vector
