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pET-28b(+) - Ecoliwiki

http://ecoliwiki.net/colipedia/index.php/pET-28b(+)

pET-28b(+)
From Ecoliwiki

At a Glance

pET-28a-c(+), image provided by Novagen (http://www.emdbiosciences.com /docs/docs/PROT/TB074.pdf)

Type

Cloning, Expression

Origin of Replication

f1 pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen (http://www.merck-chemicals.com/life-science-research /vector-table-pet-28b-plus-vector-sequence /c_HS6b.s1Oi30AAAEhncwLddSj?WFSimpleSearch_NameOrID=pET28b%28%2B%29+sequence&BackButtonText=search+results)

Features

T7 Promoter MCS His Tag lacI

Accession Number edit table (http://ecoliwiki.net/colipedia?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.P46fbbbbbca5cd&page=112641&pagename=pET-28b(%2B)&type=1&template=PlasmidTable)

Notes
The pET-28b(+) vector carries an N-terminal HisTag/thrombin/T7Tag configuration in addition to an optional C-terminal HisTag sequence. Unique sites are shown on the circle map. The T7 expression region is reversed on the circular map due to the sequence numbering from the pBR322 convention. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, the T7 terminator primer should be used when performing single stranded sequencing.

History
You can help EcoliWiki by writing about the history of this plasmid.

Sources
Novagen (http://www.emdbiosciences.com/product/69865)

References
[1] [2] [3] [4] [5]

1. Surez, T et al. (2002) Biochemical defects in retina-specific human ATP binding cassette transporter nucleotide binding domain 1 mutants associated with macular degeneration. J. Biol. Chem. 277 21759-67 PubMed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve& db=pubmed&dopt=Abstract&list_uids=11919200) Ecoliwiki page 2. Kadkhodayan, S et al. (2000) Cloning, expression, and one-step purification of the minimal essential domain of the light chain of botulinum neurotoxin type A. Protein Expr. Purif. 19 125-30 PubMed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed& dopt=Abstract&list_uids=10833399) Ecoliwiki page 3. Lecker, SH et al. (1999) Ubiquitin conjugation by the N-end rule pathway and mRNAs for its components increase in muscles of diabetic rats. J. Clin. Invest. 104 1411-20 PubMed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract& list_uids=10562303) Ecoliwiki page 4. Wang, D & Parrish, CR (1999) A heterogeneous nuclear ribonucleoprotein A/B-related protein binds to single-stranded DNA near the 5' end or within the genome of feline parvovirus and can modify virus replication. J. Virol. 73 7761-8 PubMed (http://www.ncbi.nlm.nih.gov/entrez /query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=10438866) Ecoliwiki page 5. Chaussivert, N et al. (1995) Complex interactions between yeast TFIIIB and TFIIIC. J. Biol. Chem. 270 15353-8 PubMed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=7797524) Ecoliwiki page

Sequence
1 61 121 181 241 301 361 421 481 541 601 661 721 781 841 901 961 1021 1081 1141 1201 1261 1321 1381 1441 1501 1561 1621 1681 1741 1801 1861 1921 1981 2041 2101 2161 2221 2281 atccggatat ggggttatgc tgttagcagc cgacggagct ggctgccgcg tatatctcct aattccccta gacgcatcgt acatcaccga tgggtatggt cattccttgc aggagtcgca cgcggtatgg gtaacgttat gtgaaccagg gagctgaatt attggcgttg aaatctcgcg gtcgaagcct attaactatc ccggcgttat gaagacggta ctgttagcgg tatctcactc tccggttttc gttgccaacg gttggtgcgg ccgccgttaa ttgctgcaac gtgaaaagaa gattcattaa cgcaattaat agccttcaac tatgactgtc tttcggcgag cggaatcttg cgagaagcag gtcgttgagg gatacgcgag agttcctcct tagttattgc cggatctcag cgaattcgga cggcaccagg tcttaaagtt tagtgagtcg ggccggcatc tggggaagat ggcaggcccc ggcggcggtg taagggagag catgatagcg acgatgtcgc ccagccacgt acattcccaa ccacctccag ccgatcaact gtaaagcggc cgctggatga ttcttgatgt cgcgactggg gcccattaag gcaatcaaat aacaaaccat atcagatggc atatctcggt ccaccatcaa tctctcaggg aaaccaccct tgcagctggc gtaagttagc ccagtcagct ttctttatca gaccgctttc cacgccctcg gccattatcg acccggctag cgaacgtgaa ttcagcaaaa tcagcggtgg tggtggtggt tcccgaccca ccgctgctgt aaacaaaatt tattaatttc accggcgcca cgggctcgcc gtggccgggg ctcaacggcc cgtcgagatc cccggaagag agagtatgcc ttctgcgaaa ccgcgtggca tctggccctg gggtgccagc ggtgcacaat ccaggatgcc ctctgaccag cgtggagcat ttctgtctcg tcagccgata gcaaatgctg gctgggcgca agtgggatac acaggatttt ccaggcggtg ggcgcccaat acgacaggtt tcactcatta ccttccggtg tgcaactcgt gctggagcgc ctcaagcctt ccggcatggc gctggcgggg gcgactgctg aacccctcaa cagcagccaa ggtggtgctc tttgctgtcc gatgatgatg atttctagag gcgggatcga caggtgcggt acttcgggct gactgttggg tcaacctact ccggacacca agtcaattca ggtgtctctt acgcgggaaa caacaactgg cacgcgccgt gtggtggtgt cttctcgcgc attgctgtgg acacccatca ctggtcgcat gcgcgtctgc gcggaacggg aatgagggca atgcgcgcca gacgataccg cgcctgctgg aagggcaatc acgcaaaccg tcccgactgg ggcaccggga ggcgcggggc aggacaggtg gacgatgatc cgtcactggt ggccccacgg ttgccttact ctgcaaaacg gacccgttta ctcagcttcc gagtgcggcc accagtcatg atgatggctg gggaattgtt gatctcgatc tgctggcgcc catgagcgct cgccatctcc actgggctgc tcgaatggcg gggtggtgaa atcagaccgt aagtggaagc cgggcaaaca cgcaaattgt cgatggtaga aacgcgtcag aagctgcctg acagtattat tgggtcacca gtctggctgg aaggcgactg tcgttcccac ttaccgagtc aagacagctc ggcaaaccag agctgttgcc cctctccccg aaagcgggca tctcgaccga atgactatcg ccggcagcgc ggcctgtcgc cccgccacca gtgcgcatga ggttagcaga tctgcgacct gaggccccaa tttcgggctt gcaagcttgt ctagccatat ctgcccatgg atccgctcac ctctacgccg tatatcgccg tgtttcggcg ttgcatgcac ttcctaatgc caaaaccttt tgtgaaacca ttcccgcgtg ggcgatggcg gtcgttgctg cgcggcgatt acgaagcggc tgggctgatc cactaatgtt tttctcccat gcaaatcgcg ctggcataaa gagtgccatg tgcgatgctg cgggctgcgc atgttatatc cgtggaccgc cgtctcactg cgcgttggcc gtgagcgcaa tgcccttgag tcgccgcact tctgggtcat ttgcggtatt aacgtttcgg tcgtgctcct atgaatcacc gagcaacaac

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pET-28b(+) - Ecoliwiki

http://ecoliwiki.net/colipedia/index.php/pET-28b(+)

2341 2401 2461 2521 2581 2641 2701 2761 2821 2881 2941 3001 3061 3121 3181 3241 3301 3361 3421 3481 3541 3601 3661 3721 3781 3841 3901 3961 4021 4081 4141 4201 4261 4321 4381 4441 4501 4561 4621 4681 4741 4801 4861 4921 4981 5041 5101 5161 5221 5281 5341

atgaatggtc caccattatg atctgtatta ccataccgcc aacccgtatc tcccccttac ctttatcaga acaggcagac cgcgcgtttc agcttgtctg tggcgggtgt cttaactatg taccgcacag ctgactcgct taatacggtt agcaaaaggc cccctgacga tataaagata tgccgcttac gctcacgctg acgaaccccc acccggtaag cgaggtatgt gaaggacagt gtagctcttg agcagattac ctgacgctca tctgcttaca tgctctaggc cgcgataatg ccagagttgt gtcagactaa actcctgatg ttagaagaat cggttgcatt gctcaggcgc cgtaatggct ccggattcag aaattaatag gccatcctat aaatatggta tttttctaag aataggggtt tttgttaaaa aatcggcaaa agtttggaac cgtctatcag gaggtgccgt gggaaagccg ggcgctggca gccgctacag

ttcggtttcc ttccggatct acgaagcgct agttgtttac gtgagcatcc acggaggcat agccagacat atctgtgaat ggtgatgacg taagcggatg cggggcgcag cggcatcaga atgcgtaagg gcgctcggtc atccacagaa caggaaccgt gcatcacaaa ccaggcgttt cggatacctg taggtatctc cgttcagccc acacgactta aggcggtgct atttggtatc atccggcaaa gcgcagaaaa gtggaacgaa taaacagtaa cgcgattaaa tcgggcaatc ttctgaaaca actggctgac atgcatggtt atcctgattc cgattcctgt aatcacgaat ggcctgttga tcgtcactca gttgtattga ggaactgcct ttgataatcc aattaattca ccgcgcacat ttcgcgttaa atcccttata aagagtccac ggcgatggcc aaagcactaa gcgaacgtgg agtgtagcgg ggcgcgtccc

gtgtttcgta gcatcgcagg ggcattgacc cctcacaacg tctctcgttt cagtgaccaa taacgcttct cgcttcacga gtgaaaacct ccgggagcag ccatgaccca gcagattgta agaaaatacc gttcggctgc tcaggggata aaaaaggccg aatcgacgct ccccctggaa tccgcctttc agttcggtgt gaccgctgcg tcgccactgg acagagttct tgcgctctgc caaaccaccg aaaggatctc aactcacgtt tacaaggggt ttccaacatg aggtgcgaca tggcaaaggt ggaatttatg actcaccact aggtgaaaat ttgtaattgt gaataacggt acaagtctgg tggtgatttc tgttggacga cggtgagttt tgatatgaat tgagcggata ttccccgaaa atttttgtta aatcaaaaga tattaaagaa cactacgtga atcggaaccc cgagaaagga tcacgctgcg attcgcca

aagtctggaa atgctgctgg ctgagtgatt ttccagtaac catcggtatc acaggaaaaa ggagaaactc ccacgctgat ctgacacatg acaagcccgt gtcacgtagc ctgagagtgc gcatcaggcg ggcgagcggt acgcaggaaa cgttgctggc caagtcagag gctccctcgt tcccttcggg aggtcgttcg ccttatccgg cagcagccac tgaagtggtg tgaagccagt ctggtagcgg aagaagatcc aagggatttt gttatgagcc gatgctgatt atctatcgat agcgttgcca cctcttccga gcgatccccg attgttgatg ccttttaaca ttggttgatg aaagaaatgc tcacttgata gtcggaatcg tctccttcat aaattgcagt catatttgaa agtgccacct aatcagctca atagaccgag cgtggactcc accatcaccc taaagggagc agggaagaaa cgtaaccacc

acgcggaagt ctaccctgtg tttctctggt cgggcatgtt attaccccca accgccctta aacgagctgg gagctttacc cagctcccgg cagggcgcgt gatagcggag accatatatg ctcttccgct atcagctcac gaacatgtga gtttttccat gtggcgaaac gcgctctcct aagcgtggcg ctccaagctg taactatcgt tggtaacagg gcctaactac taccttcgga tggttttttt tttgatcttt ggtcatgaac atattcaacg tatatgggta tgtatgggaa atgatgttac ccatcaagca ggaaaacagc cgctggcagt gcgatcgcgt cgagtgattt ataaactttt accttatttt cagaccgata tacagaaacg ttcatttgat tgtatttaga gaaattgtaa ttttttaacc atagggttga aacgtcaaag taatcaagtt ccccgattta gcgaaaggag acacccgccg

cagcgccctg gaacacctac cccgccgcat catcatcagt tgaacagaaa acatggcccg acgcggatga gcagctgcct agacggtcac cagcgggtgt tgtatactgg cggtgtgaaa tcctcgctca tcaaaggcgg gcaaaaggcc aggctccgcc ccgacaggac gttccgaccc ctttctcata ggctgtgtgc cttgagtcca attagcagag ggctacacta aaaagagttg gtttgcaagc tctacggggt aataaaactg ggaaacgtct taaatgggct gcccgatgcg agatgagatg ttttatccgt attccaggta gttcctgcgc atttcgtctc tgatgacgag gccattctca tgacgagggg ccaggatctt gctttttcaa gctcgatgag aaaataaaca acgttaatat aataggccga gtgttgttcc ggcgaaaaac ttttggggtc gagcttgacg cgggcgctag cgcttaatgc

Retrieved from "http://ecoliwiki.net/colipedia/index.php?title=pET-28b(%2B)&oldid=1321801"


c
Cloning Plasmids

k
Kanamycin Marker Plasmids

n
Novagen Plasmids

e l
Expression Plasmids lacI

p
Plasmids

h m
His Tag MCS

t
T7 Promoter

Category:

This page was last modified on 25 August 2011, at 13:47. Content is available under .

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