Ron Lab


 

FLAG-epitope tagged IRE1-spliced XBP-1 in the mammalian expression plasmid pFLAG-CMV-2

(see: Calfon, M., Zeng, H., Urano, F., Till, J. H., Hubbard, S. R., Harding, H. P., Clark, S. G., and Ron, D. (2002). IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA. Nature in press.)

Note: Cheah Kathryn Song Eng <hrmbdkc@hkusua.hku.hk> reported on November 4, 2004 that the sequence of this plasmid deviates from the predicted by G1028T. This impacts on the CDS changing Gly27Val. We do not know if this is a mutation that crept into our construction, a mutation that was present in the cDNA we purchased or if it is a point in which the mouse genome has some variability. Nor do we know the functional significance of this, but we note that the corresponding residue in the human is Ala. Furthermore, this variation/mutation is in a region of relatively poor sequence conservation across species, suggesting that it might not be all that important.

pFLAG.XBP1p.CMV2 (6326, predicted).

FH Key Location/Qualifiers
FH
FT CDS 925..2061
FT /note="FLAG+ORF2
FT /codon_start=1"
FT misc_difference 1429..1430
FT /note="IRE1 processing"
FT misc_feature 928..951
FT /note="FLAG epitope"
FT misc_feature 1135..1356
FT /note="bZIP"
FT misc_feature 2214..2652
FT /note="mEST: 3168224 (BE457016)"
FT misc_feature 2544..2652
FT /note="mEST 2811811, AW241077, uq39h10.x1"
FT polyA_signal 2664..3290
FT /note="hGH PolyA"
FT primer 950..973
FT /note="mXBP1.HD3.1S"
FT primer 1713..1738
FT /note="mXBP1.Xba1.2AS"
FT primer 2643..2652
FT /note="mXBP1.Xba1.2AS"
FT promoter 308..892
FT /note="CMV promoter"
XX
SQ Sequence 6326 BP; 1522 A; 1627 C; 1544 G; 1633 T;
CCCATTCGCC ATTCAGGCTG CGCAACTGTT GGGAAGGGCG ATCGGTGCGG GCCTCTTCGC TATTACGCCA GCTGGCGAAA GGGGGATGTG CTGCAAGGCG ATTAAGTTGG GTAACGCCCA GGGTTTTCCC AGTCACGACG TTGTAAAACG ACGGCCAGTG CCAAGCTGAT CTAATCAATA TTGGCCATTA GCCATATTAT TCATTGGTTA TATAGCATAA ATCAATATTG GCTATTGGCC ATTGCATACG TTGTATCCAT ATCATAATAT GTACATTTAT ATTGGCTCAT GTCCAACATT ACCGCCATGT TGACATTGAT TATTGACTAG TTATTAATAG TAATCAATTA CGGGGTCATT AGTTCATAGC CCATATATGG AGTTCCGCGT TACATAACTT ACGGTAAATG GCCCGCCTGG CGACCGCCCA GCGACCCCCG CCCGTTGACG TCAATAGTGA CGTATGTTCC CATAGTAACG CCAATAGGGA CTTTCCATTG ACGTCAATGG GTGGAGTATT TACGGTAAAC TGCCCACTTG GCAGTACATC AAGTGTATCA TATGCCAAGT CCGCCCCCTA TTGACGTCAA TGACGGTAAA TGGCCCGCCT AGCATTATGC CCAGTACATG ACCTTACGGG AGTTTCCTAC TTGGCAGTAC ATCTACGTAT TAGTCATCGC TATTACCATG GTGATGCGGT TTTGGCAGTA CACCAATGGG CGTGGATAGC GGTTTGACTC ACGGGGATTT CCAAGTCTCC ACCCCATTGA CGTCAATGGG AGTTTGTTTT GGCACCAAAA TCAACGGGAC TTTCCAAAAT GTCGTAATAA CCCCGCCCCG TTGACGCAAA TGGGCGGTAG GCGTGTACGG TGGGAGGTCT ATATAAGCAG AGCTCGTTTA GTGAACCGTC AGAATTGATC TACCATGGAC TACAAAGACG ATGACGACAA GCTTGTGGTG GCAGCGGCGC CGAGCGCGGC CACGGCGGCC CCCAAAGTGC TACTCTTATC TGGCCAGCCC GCCTCCGGCG GCCGGGCGCT GCCGCTCATG GTACCCGGTC CGCGGGCAGC AGGGTCGGAG GCGAGCGGGA CACCGCAGGC TCGCAAGCGG CAGCGGCTCA CGCACCTGAG CCCGGAGGAG AAAGCGCTGC GGAGGAAACT GAAAAACAGA GTAGCAGCGC AGACTGCTCG AGATAGAAAG AAAGCCCGGA TGAGCGAGCT GGAGCAGCAA GTGGTGGATT TGGAAGAAGA GAACCACAAA CTCCAGCTAG AAAATCAGCT TTTACGGGAG AAAACTCACG GCCTTGTGGT TGAGAACCAG GAGTTAAGAA CACGCTTGGG AATGGACACG CTGGATCCTG ACGAGGTTCC AGAGGTGGAG GCCAAGGGGA GTGGAGTAAG GCTGGTGGCC GGGTCTGCTG AGTCCGCAGC AGGTGCAGGC CCAGTTGTCA CCTCCCCAGA ACATCTTCCC ATGGACTCTG ACACTGTTGC CTCTTCAGAT TCTGAGTCTG ATATCCTTTT GGGCATTCTG GACAAGTTGG ACCCTGTCAT GTTTTTCAAA TGTCCTTCCC CAGAGTCTGC TAGTCTGGAG GAACTCCCAG AGGTCTACCC AGAAGGACCT AGTTCCTTAC CAGCCTCCCT TTCTCTGTCA GTGGGGACCT CATCAGCCAA GCTGGAAGCC ATTAATGAAC TCATTCGTTT TGACCATGTA TACACCAAGC CTCTAGTTTT AGAGATCCCC TCTGAGACAG AGAGTCAAAC TAACGTGGTA GTGAAAATTG AGGAAGCACC TCTAAGCTCT TCAGAAGAGG ATCACCCTGA ATTCATTGTC TCAGTGAAGA AAGAGCCTTT GGAAGATGAC TTCATCCCAG AGCTGGGCAT CTCAAACCTG CTTTCATCCA GCCATTGTCT GAGACCACCT TCTTGCCTGC TGGACGCTCA CAGTGACTGT GGATATGAGG GCTCCCCTTC TCCCTTCAGT GACATGTCTT CTCCACTTGG TACAGACCAC TCCTGGGAGG ATACTTTTGC CAATGAACTT TTCCCCCAGC TGATTAGTGT CTAAAGAGCC ACATAACACT GGGCCCCTTT CCCTGACCAT CACATTGCCT AGAGGATAGC ATAGGCCTGT CTCTTTCGTT AAAAGCCAAA GTAGAGGCTG TCTGGCCTTA GAAGAATTCC TCTAAAGTAT TTCAAATCTC ATAGATGACT TCCAAGTATT GTCGTTTGAC ACTCAGCTGT CTAAGGTATT CAAAGGTATT CCAGTACTAC AGCTTTTGAG ATTCTAGTTT ATCTTAAAGG TGGTAGTATA CTCTAAATCG CAGGGAGGGT CATTTGACAG TTTTTTCCCA GCCTGGCTTC AAACTATGTA GCCGAGGCTA GGCAGAAACT TCTGACCCTC TTGACCCCAC CTCCCAAGTG CTGGGCTTCA CCAGGTGTGC ACCTCCACAC CTGCCCCCCC GACATGTCAG GTGGACATGG GATTCATGAA TGGCCCTTAG CATTTCTTTC TCCACTCTCT GCTTCCCAGG TTTCGTAACC TGAGGGGGCT TGTTTTCCCT TATGTGCATT TTAAATGAAG ATCAAGAATC TTTGTAAAAT GATGAAAATT TACTATGTAA ATGCTTGATG GATCTTCTTG CTAGTGTAGC TTCTAGAGGA TCCCGGGTGG CATCCCTGTG ACCCCTCCCC AGTGCCTCTC CTGGCCTTGG AAGTTGCCAC TCCAGTGCCC ACCAGCCTTG TCCTAATAAA ATTAAGTTGC ATCATTTTGT CTGACTAGGT GTCCTCTATA ATATTATGGG GTGGAGGGGG GTGGTATGGA GCAAGGGGCC CAAGTTGGGA AGACAACCTG TAGGGCCTGC GGGGTCTATT CGGGAACCAA GCTGGAGTGC AGTGGCACAA TCTTGGCTCA CTGCAATCTC CGCCTCCTGG GTTCAAGCGA TTCTCCTGCC TCAGCCTCCC GAGTTGTTGG GATTCCAGGC ATGCATGACC AGGCTCAGCT AATTTTTGTT TTTTTGGTAG AGACGGGGTT TCACCATATT GGCCAGGCTG GTCTCCAACT CCTAATCTCA GGTGATCTAC CCACCTTGGC CTCCCAAATT GCTGGGATTA CAGGCGTGAA CCACTGCTCC CTTCCCTGTC CTTCTGATTT TAAAATAACT ATACCAGCAG GAGGACGTCC AGACACAGCA TAGGCTACCT GCCATGGCCC AACCGGTGGG ACATTTGAGT TGCTTGCTTG GCACTGTCCT CTCATGCGTT GGGTCCACTC AGTAGATGCC TGTTGAATTG GGTACGCGGC CAGCTTCTGT GGAATGTGTG TCAGTTAGGG TGTGGAAAGT CCCCAGGCTC CCCAGCAGGC AGAAGTATGC AAAGCATGCA TCTCAATTAG TCAGCAACCA GGTGTGGAAA AGTCCCCAGG CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT GCATCTCAAT TAGTCAGCAA CCATAGTCCC GCCCCTAACT CCGCCCATCC CGCCCCTAAC TCCGCCCAGT TCCGCCCATT CTCCGCCCCA TGGCTGACTA ATTTTTTTTA TTTATGCAGA GGCCGAGGCC GCCTCGGCCT CTGAGCTATT CCAGAAGTAG TGAGGAGGCT TTTTTGGAGG CCTAGGCTTT TGCAAAAAGC TCCTCGAGGA ACTGAAAAAC CAGAAAGTTA ATTCCCTATA GTGAGTCGTA TTAAATTCGT AATCATGTCA TAGCTGTTTC CTGTGTGAAA TTGTTATCCG CTCACAATTC CACACAACAT ACGAGCCGGA AGCATAAAGT GTAAAGCCTG GGGTGCCTAA TGAGTGAGCT AACTCACATT AATTGCGTTG CGCTCACTGC CCGCTTTCCA GTCGGGAAAC CTGTCGTGCC AGCTGCATTA ATGAATCGGC CAACGCGCGG GGAGAGGCGG TTTGCGTATT GGGCGCTCTT CCGCTTCCTC GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCAATGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGA ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG TGCCACCTGA CGCGCCCTGT AGCGGCGCAT TAAGCGCGGC GGGTGTGGTG GTTACGCGCA GCGTGACCGC TACACTTGCC AGCGCCCTAG CGCCCGCTCC TTTCGCTTTC TTCCCTTCCT TTCTCGCCAC GTTCGCCGGC TTTCCCCGTC AAGCTCTAAA TCGGGGCATC CCTTTAGGGT TCCGATTTAG TGCTTTACGG CACCTCGACC CCAAAAAACT TGATTAGGGT GATGGTTCAC GTAGTGGGCC ATCGCCCTGA TAGACGGTTT TTCGCCCTTT GACGTTGGAG TCCACGTTCT TTAATAGTGG ACTCTTGTTC CAAACTGGAA CAACACTCAA CCCTATCTCG GTCTATTCTT TTGATTTATA AGGGATTTTG CCGATTTCGG CCTATTGGTT AAAAAATGAG CTGATTTAAC AAAAATTTAA CGCGAATTTT AACAAAATAT TAAACGTTTA CAATTT //