cDNA Clone alpha-Tubulin HISTONE H2B

• Synonyms: TUBA1A
• Plasmid: pcDNA5D FRT/TO-GFP-TUBA1A IRES mCherry-Histone H2B
• Parent Plasmid: pcDNA5D FRT/TO IRES mCherry mod
• Expressed: GFP-TUBA1A; mCherry-Histone H2B
• Genbank: 7846 TUBA1A; M60750.1 GI:184077 Histone H2B
• Tag: GFP
• Tag: mCherry
• Species: Human

Sequence of Insert

GFP-TUBA1A cssette
GCTAGCGCCACCATGCATGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGT
GGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCA
GCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTG
AAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGT
GACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACA
TGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAG
GAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGA
GGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCA
TCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAAC
TACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCAT
CAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGC
TCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTG
CTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCC
CAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCG
GGATCACTCTCGGCATGGACGAGCTGTACAAGTCCGGACTCGGATCCATG
CGTGAGTGCATCTCCATCCACGTTGGCCAGGCTGGTGTCCAGATTGGCAA
TGCCTGCTGGGAGCTCTACTGCCTGGAACACGGCATCCAGCCCGATGGCC
AGATGCCAAGTGACAAGACCATTGGGGGAGGAGATGACTCCTTCAACACC
TTCTTCAGTGAGACGGGCGCTGGCAAGCACGTGCCCCGGGCTGTGTTTGT
AGACTTGGAACCCACAGTCATTGATGAAGTTCGCACTGGCACCTACCGCC
AGCTCTTCCACCCTGAGCAGCTCATCACAGGCAAGGAAGATGCTGCCAAT
AACTATGCCCGAGGGCACTACACCATTGGCAAGGAGATCATTGACCTTGT
GTTGGACCGAATTCGCAAGCTGGCTGACCAGTGCACCGGTCTTCAGGGCT
TCTTGGTTTTCCACAGCTTTGGTGGGGGAACTGGTTCTGGGTTCACCTCC
CTGCTCATGGAACGTCTCTCAGTTGATTATGGCAAGAAATCCAAGCTGGA
GTTCTCCATTTACCCGGCACCCCAGGTTTCCACAGCTGTAGTTGAGCCCT
ACAACTCCATCCTCACCACCCACACCACCCTGGAGCACTCTGATTGTGCC
TTCATGGTAGACAATGAGGCCATCTATGACATCTGTCGTAGAAACCTCGA
TATCGAGCGCCCAACCTACACTAACCTTAACCGCCTTATTGGTCAGATTG
TGTCCTCCATCACTGCTTCCCTGAGATTTGATGGAGCCCTGAATGTTGAC
CTGACAGAATTCCAGACCAACCTGGTGCCCTACCCCCGCATCCACTTCCC
TCTGGCCACATATGCCCCTGTCATCTCTGCTGAGAAAGCCTACCATGAAC
AGCTTTCTGTAGCAGAGATCACCAATGCTTGCTTTGAGCCAGCCAACCAG
ATGGTGAAATGTGACCCTCGCCATGGTAAATACATGGCTTGCTGCCTGTT
GTACCGTGGTGACGTGGTTCCCAAAGATGTCAATGCTGCCATTGCCACCA
TCAAAACCAAGCGCACCATCCAGTTTGTGGATTGGTGCCCCACTGGCTTC
AAGGTTGGCATCAACTACCAGCCTCCCACTGTGGTGCCTGGTGGAGACCT
GGCCAAGGTACAGAGAGCTGTGTGCATGCTGAGCAACACCACAGCCATTG
CTGAGGCCTGGGCTCGCCTGGACCACAAGTTTGACCTGATGTATGCCAAG
CGTGCCTTTGTTCACTGGTACGTGGGTGAGGGGATGGAGGAAGGCGAGTT
TTCAGAGGCCCGTGAAGATATGGCTGCCCTTGAGAAGGATTATGAGGAGG
TTGGTGTGGATTCTGTTGAAGGAGAGGGTGAGGAAGAAGGAGAGGAATAC
TAActcgag

H2B
ATCGATGGAGCTGGTGCAGGCGCTGGAGCGGGTGCCATGCCTGAACCAGC
TAAGTCAGCTCCTGCTCCGAAGAAGGGTTCCAAGAAGGCTGTGACCAAGG
CGCAGAAGAAGGATGGCAAGAAGCGCAAGCGCAGTCGTAAGGAGAGCTAC
TCCGTGTATGTGTACAAGGTGCTAAAACAGGTTCACCCCGATACTGGCAT
CTCATCCAAGGCCATGGGCATCATGAATTCCTTCGTTAACGACATCTTCG
AACGCATCGCAGGCGAGGCTTCCCGTCTGGCCCACTACAACAAGCGCTCG
ACCATTACCTCCAGGGAGATCCAGACCGCCGTGCGTCTGCTGCTTCCCGG
AGAGCTGGCCAAGCACGCAGTGTCCGAAGGTACCAAGGCTGTCACCAAGT
ATACAAGCTCCAAGTAACGCCGGCG

Amino Acid Sequence

GFP-TUBA1A cassette
MHVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFIC
TTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI
FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSH
NVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDN
HYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGLGSMRECI
SIHVGQAGVQIGNACWELYCLEHGIQPDGQMPSDKTIGGGDDSFNTFFSE
TGAGKHVPRAVFVDLEPTVIDEVRTGTYRQLFHPEQLITGKEDAANNYAR
GHYTIGKEIIDLVLDRIRKLADQCTGLQGFLVFHSFGGGTGSGFTSLLME
RLSVDYGKKSKLEFSIYPAPQVSTAVVEPYNSILTTHTTLEHSDCAFMVD
NEAIYDICRRNLDIERPTYTNLNRLIGQIVSSITASLRFDGALNVDLTEF
QTNLVPYPRIHFPLATYAPVISAEKAYHEQLSVAEITNACFEPANQMVKC
DPRHGKYMACCLLYRGDVVPKDVNAAIATIKTKRTIQFVDWCPTGFKVGI
NYQPPTVVPGGDLAKVQRAVCMLSNTTAIAEAWARLDHKFDLMYAKRAFV
HWYVGEGMEEGEFSEAREDMAALEKDYEEVGVDSVEGEGEEEGEEY*

mCherry-H2B cassette
MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWER
VMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEA
SSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNV
NIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKSGLIDGAGAGAGAG
AMPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSVYVYKVLKQV
HPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAV
RLLLPGELAKHAVSEGTKAVTKYTSSK*

• Vector Type: Mammalian
• Antibiotic: Amp
• Full Snapgene Map: pcDNA5D FRT/TO-GFP-TUBA1A IRES mCherry-Histone H2B63.67 KBSnapgene files can be opened using software available here: www.snapgene.com
• DU Number: DU28877
• Unit Source: Protein Phosphorylation and Ubiquitylation Unit
• Location: PPU
• MTA: Dundee
• Notes: Parental modified IRES-mCherry containing GFP-TUBA1A inserted upstream of IRES-mCherry (28861). TUBA1A contains the following silent changes compared to NCBI ref shown:; D47 (GAT > GAC), G57 (GGG > GGC), H61 (CAT > CAC), A65 (GCA > GCT), Q91 (CAA > CAG), L92 (CTT > CTC), K96 (AAA > AAG), L117 (CTC > CTT), A126 (GCC > GCT), T130 (ACG > ACC), L132 (CTC > CTT), S151 (TCG > TCC), K164 (AAG > AAA), S170 (TCT > TCC), A174 (GCG > GCA), I219 (ATT > ATC), R221 (CGT > CGC), Y224 (TAT > TAC), L227 (CTG > CTT), N228 (AAT > AAC), R229 (AGG > CGC), L230 (TTA > CTT), I231 (ATA > ATT), Q233 (CAA > CAG), Y262 (TAT > TAC), K336 (AAG > AAA), R339 (CGT > CGC), K401 (AAA > AAG), V409 (GTT > GTG), G410 (GGG > GGT), G416 (GGT > GGC), E423 (GAG > GAA), D424 (GAC > GAT),
• Low Temp: No