Ron Lab


 

FLAG-epitope tagged unspliced 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.XBP1u.CMV2 (6352, predicted).

FH Key Location/Qualifiers
FH
FT promoter 308..892
FT /note="CMV promoter"
FT CDS 925..1749
FT /note="FLAG+ORF1
FT /codon_start=1"
FT misc_feature 928..951
FT /note="FLAG epitope"
FT primer 950..973
FT /note="mXBP1.HD3.1S"
FT misc_feature 1135..1356
FT /note="bZIP"
FT misc_feature 1430..1455
FT /note="IRE1 processing"
FT primer 1739..1764
FT /note="mXBP1.Xba1.2AS"
FT misc_feature 2240..2678
FT /note="mEST: 3168224 (BE457016)"
FT misc_feature 2570..2678
FT /note="mEST 2811811, AW241077, uq39h10.x1"
FT primer 2669..2678
FT /note="mXBP1.Xba1.2AS"
FT polyA_signal 2690..3316
FT /note="hGH PolyA"
XX
SQ Sequence 6352 BP; 1528 A; 1636 C; 1549 G; 1639 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 ACTCAGACTA TGTGCACCTC TGCAGCAGGT GCAGGCCCAG TTGTCACCTC CCCAGAACAT CTTCCCATGG ACTCTGACAC TGTTGCCTCT TCAGATTCTG AGTCTGATAT CCTTTTGGGC ATTCTGGACA AGTTGGACCC TGTCATGTTT TTCAAATGTC CTTCCCCAGA GTCTGCTAGT CTGGAGGAAC TCCCAGAGGT CTACCCAGAA GGACCTAGTT CCTTACCAGC CTCCCTTTCT CTGTCAGTGG GGACCTCATC AGCCAAGCTG GAAGCCATTA ATGAACTCAT TCGTTTTGAC CATGTATACA CCAAGCCTCT AGTTTTAGAG ATCCCCTCTG AGACAGAGAG TCAAACTAAC GTGGTAGTGA AAATTGAGGA AGCACCTCTA AGCTCTTCAG AAGAGGATCA CCCTGAATTC ATTGTCTCAG TGAAGAAAGA GCCTTTGGAA GATGACTTCA TCCCAGAGCT GGGCATCTCA AACCTGCTTT CATCCAGCCA TTGTCTGAGA CCACCTTCTT GCCTGCTGGA CGCTCACAGT GACTGTGGAT ATGAGGGCTC CCCTTCTCCC TTCAGTGACA TGTCTTCTCC ACTTGGTACA GACCACTCCT GGGAGGATAC TTTTGCCAAT GAACTTTTCC CCCAGCTGAT TAGTGTCTAA AGAGCCACAT AACACTGGGC CCCTTTCCCT GACCATCACA TTGCCTAGAG GATAGCATAG GCCTGTCTCT TTCGTTAAAA GCCAAAGTAG AGGCTGTCTG GCCTTAGAAG AATTCCTCTA AAGTATTTCA AATCTCATAG ATGACTTCCA AGTATTGTCG TTTGACACTC AGCTGTCTAA GGTATTCAAA GGTATTCCAG TACTACAGCT TTTGAGATTC TAGTTTATCT TAAAGGTGGT AGTATACTCT AAATCGCAGG GAGGGTCATT TGACAGTTTT TTCCCAGCCT GGCTTCAAAC TATGTAGCCG AGGCTAGGCA GAAACTTCTG ACCCTCTTGA CCCCACCTCC CAAGTGCTGG GCTTCACCAG GTGTGCACCT CCACACCTGC CCCCCCGACA TGTCAGGTGG ACATGGGATT CATGAATGGC CCTTAGCATT TCTTTCTCCA CTCTCTGCTT CCCAGGTTTC GTAACCTGAG GGGGCTTGTT TTCCCTTATG TGCATTTTAA ATGAAGATCA AGAATCTTTG TAAAATGATG AAAATTTACT ATGTAAATGC TTGATGGATC TTCTTGCTAG TGTAGCTTCT AGAGGATCCC GGGTGGCATC CCTGTGACCC CTCCCCAGTG CCTCTCCTGG CCTTGGAAGT TGCCACTCCA GTGCCCACCA GCCTTGTCCT AATAAAATTA AGTTGCATCA TTTTGTCTGA CTAGGTGTCC TCTATAATAT TATGGGGTGG AGGGGGGTGG TATGGAGCAA GGGGCCCAAG TTGGGAAGAC AACCTGTAGG GCCTGCGGGG TCTATTCGGG AACCAAGCTG GAGTGCAGTG GCACAATCTT GGCTCACTGC AATCTCCGCC TCCTGGGTTC AAGCGATTCT CCTGCCTCAG CCTCCCGAGT TGTTGGGATT CCAGGCATGC ATGACCAGGC TCAGCTAATT TTTGTTTTTT TGGTAGAGAC GGGGTTTCAC CATATTGGCC AGGCTGGTCT CCAACTCCTA ATCTCAGGTG ATCTACCCAC CTTGGCCTCC CAAATTGCTG GGATTACAGG CGTGAACCAC TGCTCCCTTC CCTGTCCTTC TGATTTTAAA ATAACTATAC CAGCAGGAGG ACGTCCAGAC ACAGCATAGG CTACCTGCCA TGGCCCAACC GGTGGGACAT TTGAGTTGCT TGCTTGGCAC TGTCCTCTCA TGCGTTGGGT CCACTCAGTA GATGCCTGTT GAATTGGGTA CGCGGCCAGC TTCTGTGGAA TGTGTGTCAG TTAGGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAG CATGCATCTC AATTAGTCAG CAACCAGGTG TGGAAAAGTC CCCAGGCTCC CCAGCAGGCA GAAGTATGCA AAGCATGCAT CTCAATTAGT CAGCAACCAT AGTCCCGCCC CTAACTCCGC CCATCCCGCC CCTAACTCCG CCCAGTTCCG CCCATTCTCC GCCCCATGGC TGACTAATTT TTTTTATTTA TGCAGAGGCC GAGGCCGCCT CGGCCTCTGA GCTATTCCAG AAGTAGTGAG GAGGCTTTTT TGGAGGCCTA GGCTTTTGCA AAAAGCTCCT CGAGGAACTG AAAAACCAGA AAGTTAATTC CCTATAGTGA GTCGTATTAA ATTCGTAATC ATGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA CAACATACGA GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT CACATTAATT GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAA TGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT GCCATTGCTA CAGGCATCGT GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC CCGGCGTCAA TACGGGATAA TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC ACCTGACGCG CCCTGTAGCG GCGCATTAAG CGCGGCGGGT GTGGTGGTTA CGCGCAGCGT GACCGCTACA CTTGCCAGCG CCCTAGCGCC CGCTCCTTTC GCTTTCTTCC CTTCCTTTCT CGCCACGTTC GCCGGCTTTC CCCGTCAAGC TCTAAATCGG GGCATCCCTT TAGGGTTCCG ATTTAGTGCT TTACGGCACC TCGACCCCAA AAAACTTGAT TAGGGTGATG GTTCACGTAG TGGGCCATCG CCCTGATAGA CGGTTTTTCG CCCTTTGACG TTGGAGTCCA CGTTCTTTAA TAGTGGACTC TTGTTCCAAA CTGGAACAAC ACTCAACCCT ATCTCGGTCT ATTCTTTTGA TTTATAAGGG ATTTTGCCGA TTTCGGCCTA TTGGTTAAAA AATGAGCTGA TTTAACAAAA ATTTAACGCG AATTTTAACA AAATATTAAA CGTTTACAAT TT //