pPIC9K_A2AR_316_V229C_R291A

246917 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, R291A
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