pET-28b(+)
At a Glance
<protect>
Type |
Cloning, Expression |
---|---|
Origin of Replication |
f1 |
Copy # |
~40 |
Markers |
Kanamycin |
Link to Sequence | |
Features |
T7 Promoter |
Accession Number |
|
edit table |
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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
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Sources
References
- ↑ 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
- ↑ 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
- ↑ 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
- ↑ 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
- ↑ Chaussivert, N et al. (1995) Complex interactions between yeast TFIIIB and TFIIIC. J. Biol. Chem. 270 15353-8 PubMed
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