pMT1219

228301 Synthetic Yeast Expression

Plasmid Information

Purpose
2,4-Diacetylphloroglucinol (DAPG)-regulatable expression of SARS-CoV-2 RBD omicron variant
Depositor
Jun Ishii
Article
Tominaga et al Nat Commun. 2024 Dec 19;15(1):10653. doi
Insert
MFalpha(2xAdv)-SARS CoV-2 receptor binding domain (RBD)-His (S Severe acute respiratory syndrome-related coronavirus 2, Synthetic)
Tags
GGG linker followed by His6 and MF_ secretion sig…
Mutation
G339D, S371L, S373P, S375F, K417N, N440K, G446S, …
Promoter
2,4-Diacetylphloroglucinol-regulatable synthetic …
Available Since
March 28, 2025
Availability
Available

Sequence Information

Length
6,892 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
AmpR bla beta-lactamase 200..1061 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
alpha-factor secretion signal MF-alpha-1 N-terminal secretion signal from S. cer… 3645..3900 Fwd
DNA Sequence
ATGAGATTTCCTTCAATTTTTACTGCAGTTTTATTCGCAGCATCCTCCGCATTAGCTGCTCCAGTCAACACTACAACAGAAGATGAAACGGCACAAATTCCGGCTGAAGCTGTCATCGGTTACTCAGATTTAGAAGGGGATTTCGATGCTGCTGTTTTGCCATTTTCCAACAGCACAAATAACGGGTTATTGTTTATAAATACTACTATTGCCAGCATTGCTGCTAAAGAAGAAGGGGTATCTTTGGATAAAAGA
Protein Sequence
MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLEGDFDAAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKR
6xHis — 6xHis affinity tag 4527..4545 Fwd
DNA Sequence
CATCATCATCACCATCAC
Protein Sequence
HHHHHH
HygR aph(4)-Ia aminoglycoside phosphotransferase from … 5265..6288 Fwd
DNA Sequence
ATGAAGAAGCCAGAGTTGACTGCTACTTCCGTTGAGAAGTTCTTGATCGAGAAGTTCGACTCCGTTTCCGACTTGATGCAATTGTCTGAAGGTGAAGAGTCCAGAGCTTTCTCCTTTGACGTTGGTGGTAGAGGTTACGTTTTGAGAGTTAATTCCTGTGCTGACGGTTTCTACAAGGACAGATACGTTTACAGACACTTCGCTTCCGCTGCTTTGCCAATTCCAGAGGTTTTGGACATTGGTGAGTTCTCCGAGTCCTTGACTTACTGTATCTCCAGAAGAGCTCAGGGAGTTACATTGCAAGACTTGCCAGAAACTGAGTTGCCAGCTGTTTTGCAACCAGTTGCTGAAGCTATGGACGCTATTGCTGCTGCTGACTTGTCTCAAACTTCTGGTTTTGGTCCATTCGGTCCACAGGGTATTGGTCAGTACACTACTTGGAGAGACTTCATCTGTGCTATCGCTGACCCACATGTTTACCACTGGCAAACTGTTATGGACGATACTGTTTCCGCTTCCGTTGCTCAGGCTTTGGACGAATTGATGTTGTGGGCTGAAGATTGTCCTGAGGTTAGACATTTGGTTCACGCTGACTTCGGTTCCAACAACGTTTTGACTGACAACGGTAGAATCACTGCTGTTATCGACTGGTCCGAAGCTATGTTCGGTGACTCTCAATACGAGGTTGCTAACATCTTCTTTTGGAGACCATGGTTGGCTTGTATGGAACAGCAGACTAGATACTTCGAGAGAAGACACCCAGAATTGGCTGGTTCCCCAAGATTGAGAGCTTACATGTTGAGAATCGGTTTGGACCAGTTGTACCAGTCCTTGGTTGATGGTAACTTCGATGATGCTGCTTGGGCTCAAGGTAGATGTGACGCTATCGTTAGATCCGGTGCTGGTACTGTTGGTAGAACTCAGATTGCTAGAAGATCCGCTGCTGTTTGGACTGACGGTTGTGTTGAAGTTTTGGCTGACTCCGGTAACAGAAGACCATCCACTAGACCAAGAGCTAAAGAG
Protein Sequence
MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
promoter 2
Label Gene Product Position Strand
AmpR promoter bla — 95..200 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lac promoter — — 2143..2174 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 2
Label Gene Product Position Strand
CAP binding site — — 2107..2129 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator — — 2181..2198 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
rep_origin 1
Label Gene Product Position Strand
ori — — 1231..1820 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTTTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
AOX1 terminator Pichia pastoris AOX1 — 4622..4869 Fwd
DNA Sequence
TCAAGAGGATGTCAGAATGCCATTTGCCTGAGAGATGCAGGCTTCATTTTTGATACTTTTTTATTTGTAACCTATATAGTATAGGATTTTTTTTGTCATTTTGTTTCTTCTCGTACGAGCTTGCTCCTGATCAGCCTATCTCGCAGCTGATGAATATCTTGTGGTAGGGGTTTGGGAAAATCATTCGAGTTTGATGTTTTTCTTGGTATTTCCCACTCCTCTTCAGAGTACAGAAGATTAAGTGAGA