PortEco logo

pET-28b(+)

From EcoliWiki
Jump to: navigation, search

At a Glance

pET-28a-c(+), image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 Promoter
MCS
His Tag
lacI

Accession Number




Notes

The pET-28b(+) vector carries an N-terminal His•Tag/thrombin/T7•Tag configuration in addition to an optional C-terminal His•Tag 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

References

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

  1. Suárez, 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 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 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 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 EcoliWiki page
  5. Chaussivert, N et al. (1995) Complex interactions between yeast TFIIIB and TFIIIC. J. Biol. Chem. 270 15353-8 PubMed EcoliWiki page

Sequence

       1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 
      61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 
     121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt 
     181 cgacggagct cgaattcgga tcccgaccca tttgctgtcc accagtcatg ctagccatat 
     241 ggctgccgcg cggcaccagg ccgctgctgt gatgatgatg atgatggctg ctgcccatgg 
     301 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac 
     361 aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg 
     421 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg 
     481 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg 
     541 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac 
     601 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc 
     661 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt 
     721 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca 
     781 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg 
     841 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg 
     901 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg 
     961 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt 
    1021 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc 
    1081 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc 
    1141 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt 
    1201 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat 
    1261 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg 
    1321 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa 
    1381 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg 
    1441 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg 
    1501 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc 
    1561 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc 
    1621 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc 
    1681 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg 
    1741 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc 
    1801 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 
    1861 cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag 
    1921 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact 
    1981 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat 
    2041 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt 
    2101 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg 
    2161 cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct 
    2221 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc 
    2281 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac 
    2341 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg 
    2401 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac 
    2461 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat 
    2521 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt 
    2581 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa 
    2641 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg 
    2701 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga 
    2761 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct 
    2821 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 
    2881 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 
    2941 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg 
    3001 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa 
    3061 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca 
    3121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 
    3181 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 
    3241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 
    3301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 
    3361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 
    3421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 
    3481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 
    3541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 
    3601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 
    3661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 
    3721 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 
    3781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 
    3841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 
    3901 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg 
    3961 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct 
    4021 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct 
    4081 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg 
    4141 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg 
    4201 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt 
    4261 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta 
    4321 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc 
    4381 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc 
    4441 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag 
    4501 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca 
    4561 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg 
    4621 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt 
    4681 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa 
    4741 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag 
    4801 tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca 
    4861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat 
    4921 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 
    4981 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 
    5041 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 
    5101 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 
    5161 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 
    5221 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 
    5281 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 
    5341 gccgctacag ggcgcgtccc attcgcca