pET15b p97-Strep-His E314Amb

169021 Homo sapiens Bacterial Expression

Plasmid Information

Purpose
Expresses p97-Strep-His E314Amb (incorporates noncanonical amino acid at position 314 via amber suppression) in bacteria
Depositor
Hemmo Meyer
Article
Weith et al Mol Cell. 2018 Oct 11. pii: S1097-2765(1
Insert
p97
Tags
His and SBP
Mutation
E314TAG
Promoter
T7
Available Since
July 7, 2021
Availability
Available

Sequence Information

Length
8,197 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
AmpR bla beta-lactamase 208..1069 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGTGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
rop rop Rop protein, which maintains plasmids a… 2254..2446 Rev
DNA Sequence
GTGACCAAATAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGTGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTK*EKTALNMARFIRSQTLTLLEKLNELDVDEQADICESLHDHADELYRSCLARFGDDGENL
SBP — streptavidin-binding peptide 7738..7852 Fwd
DNA Sequence
ATGGACGAGAAGACAACGGGATGGCGTGGAGGTCACGTAGTGGAGGGGCTTGCGGGTGAATTGGAGCAATTACGCGCCCGCCTTGAGCATCACCCGCAGGGCCAGCGGGAGCCC
Protein Sequence
MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP
6xHis — 6xHis affinity tag 7852..7870 Fwd
DNA Sequence
CACCATCACCATCACCAT
Protein Sequence
HHHHHH
promoter 4
Label Gene Product Position Strand
AmpR promoter bla — 103..208 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lacI promoter lacI — 4839..4917 Rev
DNA Sequence
GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
T7 promoter — — 5225..5244 Fwd
DNA Sequence
TAATACGACTCACTATAGG
tet promoter — — 8159..8188 Rev
DNA Sequence
TTGACAGCTTATCATCGATAAGCTTTAAT
protein_bind 1
Label Gene Product Position Strand
lac operator — — 5244..5269 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
rep_origin 1
Label Gene Product Position Strand
ori — — 1239..1828 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
T7 terminator — — 7937..7985 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG