pPIC9K_A2AR_316_V229C_R293A

246918 Homo sapiens Yeast Expression

Plasmid Information

Purpose
Express A2AR mutants in Pichia Pastoris
Depositor
Libin Ye
Article
Wang et al Nat Commun. 2023 Mar 10;14(1):1325. doi:
Insert
Adenosine A2A receptor (ADORA2A Human)
Tags
10xHis and alpha-factor secretion signal, FLAG
Mutation
truncated to aa 1-316, V229C, R293A
Promoter
AOX1
Available Since
Nov. 5, 2025
Availability
Available

Sequence Information

Length
10,303 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 7
Label Gene Product Position Strand
alpha-factor secretion signal MF-alpha-1 N-terminal secretion signal from S. cer… 948..1203 Fwd
DNA Sequence
ATGAGATTTCCTTCAATTTTTACTGCAGTTTTATTCGCAGCATCCTCCGCATTAGCTGCTCCAGTCAACACTACAACAGAAGATGAAACGGCACAAATTCCGGCTGAAGCTGTCATCGGTTACTCAGATTTAGAAGGGGATTTCGATGTTGCTGTTTTGCCATTTTCCAACAGCACAAATAACGGGTTATTGTTTATAAATACTACTATTGCCAGCATTGCTGCTAAAGAAGAAGGGGTATCTCTCGAAAAAAGA
Protein Sequence
MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLEGDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKR
FLAG — FLAG(R) epitope tag, followed by an ent… 1203..1227 Fwd
DNA Sequence
GACTACAAGGACGATGACGATAAG
Protein Sequence
DYKDDDDK
TEV site — tobacco etch virus (TEV) protease recog… 1266..1287 Fwd
DNA Sequence
GAAAACTTGTATTTCCAGGGC
Protein Sequence
ENLYFQG
10xHis — 10xHis affinity tag 2235..2265 Fwd
DNA Sequence
CATCACCATCACCATCACCATCACCATCAC
Protein Sequence
HHHHHHHHHH
PpHIS4 Pichia pastoris HIS4 multifunctional enzyme, required for hi… 3014..5549 Rev
DNA Sequence
ATGACATTTCCCTTGCTACCTGCATACGCAAGTGTTGCAGAGTTTGATAATTCCTTGAGTTTGGTAGGAAAAGCCGTGTTTCCCTATGCTGCTGACCAGCTGCACAACCTGATCAAGTTCACTCAATCGACTGAGCTTCAAGTTAATGTGCAAGTTGAGTCATCCGTTACAGAGGACCAATTTGAGGAGCTGATCGACAACTTGCTCAAGTTGTACAATAATGGTATCAATGAAGTGATTTTGGACCTAGATTTGGCAGAAAGAGTTGTCCAAAGGATGATCCCAGGCGCTAGGGTTATCTATAGGACCCTGGTTGATAAAGTTGCATCCTTGCCCGCTAATGCTAGTATCGCTGTGCCTTTTTCTTCTCCACTGGGCGATTTGAAAAGTTTCACTAATGGCGGTAGTAGAACTGTTTATGCTTTTTCTGAGACCGCAAAGTTGGTAGATGTGACTTCCACTGTTGCTTCTGGTATAATCCCCATTATTGATGCTCGGCAATTGACTACTGAATACGAACTTTCTGAAGATGTCAAAAAGTTCCCTGTCAGTGAAATTTTGTTGGCGTCTTTGACTACTGACCGCCCCGATGGTCTATTCACTACTTTGGTGGCTGACTCTTCTAATTACTCGTTGGGCCTGGTGTACTCGTCCAAAAAGTCTATTCCGGAGGCTATAAGGACACAAACTGGAGTCTACCAATCTCGTCGTCACGGTTTGTGGTATAAAGGTGCTACATCTGGAGCAACTCAAAAGTTGCTGGGTATCGAATTGGATTGTGATGGAGACTGCTTGAAATTTGTGGTTGAACAAACAGGTGTTGGTTTCTGTCACTTGGAACGCACTTCCTGTTTTGGCCAATCAAAGGGTCTTAGAGCCATGGAAGCCACCTTGTGGGATCGTAAGAGCAATGCTCCAGAAGGTTCTTATACCAAACGGTTATTTGACGACGAAGTTTTGTTGAACGCTAAAATTAGGGAGGAAGCTGATGAACTTGCAGAAGCTAAATCCAAGGAAGATATAGCCTGGGAATGTGCTGACTTATTTTATTTTGCATTAGTTAGATGTGCCAAGTACGGTGTGACGTTGGACGAGGTGGAGAGAAACCTGGATATGAAGTCCCTAAAGGTCACTAGAAGGAAAGGAGATGCCAAGCCAGGATACACCAAGGAACAACCTAAAGAAGAATCCAAACCTAAAGAAGTCCCTTCTGAAGGTCGTATTGAATTGTGCAAAATTGACGTTTCTAAGGCCTCCTCACAAGAAATTGAAGATGCCCTTCGTCGTCCTATCCAGAAAACGGAACAGATTATGGAATTAGTCAAACCAATTGTCGACAATGTTCGTCAAAATGGTGACAAAGCCCTTTTAGAACTAACTGCCAAGTTTGATGGAGTCGCTTTGAAGACACCTGTGTTAGAAGCTCCTTTCCCAGAGGAACTTATGCAATTGCCAGATAACGTTAAGAGAGCCATTGATCTCTCTATAGATAACGTCAGGAAATTCCATGAAGCTCAACTAACGGAGACGTTGCAAGTTGAGACTTGCCCTGGTGTAGTCTGCTCTCGTTTTGCAAGACCTATTGAGAAAGTTGGCCTCTATATTCCTGGTGGAACCGCAATTCTGCCTTCCACTTCCCTGATGCTGGGTGTTCCTGCCAAAGTTGCTGGTTGCAAAGAAATTGTTTTTGCATCTCCACCTAAGAAGGATGGTACCCTTACCCCAGAAGTCATCTACGTTGCCCACAAGGTTGGTGCTAAGTGTATCGTGCTAGCAGGAGGCGCCCAGGCAGTAGCTGCTATGGCTTACGGAACAGAAACTGTTCCTAAGTGTGACAAAATATTTGGTCCAGGAAACCAGTTCGTTACTGCTGCCAAGATGATGGTTCAAAATGACACATCAGCCCTGTGTAGTATTGACATGCCTGCTGGGCCTTCTGAAGTTCTAGTTATTGCTGATAAATACGCTGATCCAGATTTCGTTGCCTCAGACCTTCTGTCTCAAGCTGAACATGGTATTGATTCCCAGGTGATTCTGTTGGCTGTCGATATGACAGACAAGGAGCTTGCCAGAATTGAAGATGCTGTTCACAACCAAGCTGTGCAGTTGCCAAGGGTTGAAATTGTACGCAAGTGTATTGCACACTCTACAACCCTATCGGTTGCAACCTACGAGCAGGCTTTGGAAATGTCCAATCAGTACGCTCCTGAACACTTGATCCTGCAAATCGAGAATGCTTCTTCTTATGTTGATCAAGTACAACACGCTGGATCTGTGTTTGTTGGTGCCTACTCTCCAGAGAGTTGTGGAGATTACTCCTCCGGTACCAACCACACTTTGCCAACGTACGGATATGCCCGTCAATACAGCGGAGTTAACACTGCAACCTTCCAGAAGTTCATCACTTCACAAGACGTAACTCCTGAGGGACTGAAACATATTGGCCAAGCAGTGATGGATCTGGCTGCTGTTGAAGGTCTAGATGCTCACCGCAATGCTGTTAAGGTTCGTATGGAGAAACTGGGACTTATTTAA
Protein Sequence
MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTELQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTLVDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPIIDARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSIPEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLERTSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKEDIAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKPKEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLELTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCPGVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPEVIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSALCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIEDAVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQVQHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLKHIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI
KanR aph(3')-Ia aminoglycoside phosphotransferase 5961..6777 Rev
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCTTGCTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
AmpR bla beta-lactamase 9132..9993 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 2
Label Gene Product Position Strand
AOX1 promoter Pichia pastoris AOX1 — 1..937 Fwd
DNA Sequence
GATCTAACATCCAAAGACGAAAGGTTGAATGAAACCTTTTTGCCATCCGACATCCACAGGTCCATTCTCACACATAAGTGCCAAACGCAACAGGAGGGGATACACTAGCAGCAGACCGTTGCAAACGCAGGACCTCCACTCCTCTTCTCCTCAACACCCACTTTTGCCATCGAAAAACCAGCCCAGTTATTGGGCTTGATTGGAGCTCGCTCATTCCAATTCCTTCTATTAGGCTACTAACACCATGACTTTATTAGCCTGTCTATCCTGGCCCCCCTGGCGAGGTTCATGTTTGTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGATGAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCAAAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGGCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTTGATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAA
AmpR promoter bla — 9993..10098 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rep_origin 1
Label Gene Product Position Strand
ori — — 8373..8962 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
AOX1 terminator Pichia pastoris AOX1 — 2355..2602 Fwd
DNA Sequence
TCAAGAGGATGTCAGAATGCCATTTGCCTGAGAGATGCAGGCTTCATTTTTGATACTTTTTTATTTGTAACCTATATAGTATAGGATTTTTTTTGTCATTTTGTTTCTTCTCGTACGAGCTTGCTCCTGATCAGCCTATCTCGCAGCTGATGAATATCTTGTGGTAGGGGTTTGGGAAAATCATTCGAGTTTGATGTTTTTCTTGGTATTTCCCACTCCTCTTCAGAGTACAGAAGATTAAGTGAGA