This page was created for an undergraduate assignment at Davidson College.
More Information on the pQE30UA expression plasmid
Click here to go to the Qiagen home page: www.qiagen.com
Click here to go to the QIAexpressionist handbook: http://www.qiagen.com/literature/handbooks/qxp/qxp/1016852QXPHB_0301WW.pdf
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-----T5 promoter, lac operon
The T5 viral promoter allows for mass production of the protein. The lac operon allows transcription to be regulated using IPTG.
----- translational start codon
----- 6x histidine epitope tag
The tag will enable us to visulaize the transcribed protein using antibodies.
----- multiple
cloning sites (MCS)
----- PCR cloning site
^ location of restriction enzyme cut
The three restriction sites identified will be used to identify the orientation of the five IDH genes.
For more information on the experiment that will determine the orientation of the insert: Determining Orientation
length: 3504 bp
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
EcoRV 121 ggatcgcatc accatcacca tcacggatcc cacgtgat^at cctcaatcgc ttcTAgaagc
BglII 181 gattgagga^g atctgagctc ggtacccggg tcgacaggcc tctgcagcca agcttaatta
241 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
301 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc
361 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
421 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
481 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
541 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
601 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
661 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
721 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
781 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
841 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
901 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
961 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
1021 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
1081 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
1141 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
1201 cgccctctag agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
1261 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
1321 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
1381 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
1441 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
1501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
1561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
1621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
1681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
1741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
1801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
1861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
1921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
1981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
2041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
2101 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
2161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
2221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
2281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
2341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
2401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
2461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
2521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
2581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
2641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
2701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
2761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
2821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
2881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
2941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
3001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
3061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
3121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
3181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
3241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
3301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
3361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
3421 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
3481 atcacgaggc cctttcgtct tcac