P1K-3HA-Atg8∆R-MET15

Plasmid Information

Purpose

Expression of 3HA-Atg8∆R. This variant bypasses the need of Atg4 for initial C-terminal processing.

Depositor

Zhiping Xie

Article

He et al BMC Biol. 2021 Jun 4;19(1):117. doi: 10.

Insert

ATG8

Sequence Information

Length

6,302 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 3

Label	Gene	Product	Position	Strand
3xHA	—	three tandem HA epitope tags	1086..1176	Fwd
DNA Sequence TACCCATACGATGTTCCTGACTATGCGGGCTATCCCTATGACGTCCCGGACTATGCAGGATCCTATCCATATGACGTTCCAGATTACGCT Protein Sequence YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA
MET17	S. cerevisiae MET17	O-acetylserine and O-acetylhomoserine s…	2014..3349	Rev
DNA Sequence ATGCCATCTCATTTCGATACTGTTCAACTACACGCCGGCCAAGAGAACCCTGGTGACAATGCTCACAGATCCAGAGCTGTACCAATTTACGCCACCACTTCTTATGTTTTCGAAAACTCTAAGCATGGTTCGCAATTGTTTGGTCTAGAAGTTCCAGGTTACGTCTATTCCCGTTTCCAAAACCCAACCAGTAATGTTTTGGAAGAAAGAATTGCTGCTTTAGAAGGTGGTGCTGCTGCTTTGGCTGTTTCCTCCGGTCAAGCCGCTCAAACCCTTGCCATCCAAGGTTTGGCACACACTGGTGACAACATCGTTTCCACTTCTTACTTATACGGTGGTACTTATAACCAGTTCAAAATCTCGTTCAAAAGATTTGGTATCGAGGCTAGATTTGTTGAAGGTGACAATCCAGAAGAATTCGAAAAGGTCTTTGATGAAAGAACCAAGGCTGTTTATTTGGAAACCATTGGTAATCCAAAGTACAATGTTCCGGATTTTGAAAAAATTGTTGCAATTGCTCACAAACACGGTATTCCAGTTGTCGTTGACAACACATTTGGTGCCGGTGGTTACTTCTGTCAGCCAATTAAATACGGTGCTGATATTGTAACACATTCTGCTACCAAATGGATTGGTGGTCATGGTACTACTATCGGTGGTATTATTGTTGACTCTGGTAAGTTCCCATGGAAGGACTACCCAGAAAAGTTCCCTCAATTCTCTCAACCTGCCGAAGGATATCACGGTACTATCTACAATGAAGCCTACGGTAACTTGGCATACATCGTTCATGTTAGAACTGAACTATTAAGAGATTTGGGTCCATTGATGAACCCATTTGCCTCTTTCTTGCTACTACAAGGTGTTGAAACATTATCTTTGAGAGCTGAAAGACACGGTGAAAATGCATTGAAGTTAGCCAAATGGTTAGAACAATCCCCATACGTATCTTGGGTTTCATACCCTGGTTTAGCATCTCATTCTCATCATGAAAATGCTAAGAAGTATCTATCTAACGGTTTCGGTGGTGTCTTATCTTTCGGTGTAAAAGACTTACCAAATGCCGACAAGGAAACTGACCCATTCAAACTTTCTGGTGCTCAAGTTGTTGACAATTTAAAGCTTGCCTCTAACTTGGCCAATGTTGGTGATGCCAAGACCTTAGTCATTGCTCCATACTTCACTACCCACAAACAATTAAATGACAAAGAAAAGTTGGCATCTGGTGTTACCAAGGACTTAATTCGTGTCTCTGTTGGTATCGAATTTATTGATGACATTATTGCAGACTTCCAGCAATCTTTTGAAACTGTTTTCGCTGGCCAAAAACCATGA Protein Sequence MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQLFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNIVSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNVPDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGIIVDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFASFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNGFGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQLNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP
AmpR	bla	beta-lactamase	4974..5835	Rev
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

promoter 4

Label	Gene	Product	Position	Strand
SP6 promoter	—	—	0..6302	Fwd
DNA Sequence ATTTAGGTGACACTATAGA
MET17 promoter	S. cerevisiae MET17	—	3349..3700	Rev
DNA Sequence TTATTTTTTGCTTTTTCTCTTGAGGTCACATGATCGCAAAATGGCAAATGGCACGTGAAGCTGTCGATATTGGGGAACTGTGGTGGTTGGCAAATGACTAATTAAGTTAGTCAAGGCGCCATCCTCATGAAAACTGTGTAACATAATAACCGAAGTGTCGAAAAGGTGGCACCTTGTCCAATTGAACACGCTCGATGAAAAAAATAAGATATATATAAGGTTAAGTAAAGCGTCTGTTAGAAAGGAAGTTTTTCCTTTTTCTTGCTCTCTTGTCTTTTCATCTACTATTTCCTTCGTGTAATACAGGGTCGTCAGATACATAGATACAATTCTATTACCCCCATCCATACA
T7 promoter	—	—	3939..3958	Rev
DNA Sequence TAATACGACTCACTATAGG
AmpR promoter	bla	—	5835..5940	Rev
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

protein_bind 2

Label	Gene	Product	Position	Strand
loxP	—	—	1798..1832	Fwd
DNA Sequence ATAACTTCGTATAATGTATGCTATACGAAGTTAT
loxP	—	—	3852..3886	Fwd
DNA Sequence ATAACTTCGTATAATGTATGCTATACGAAGTTAT

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	4215..4804	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
ADH1 terminator	S. cerevisiae ADH1	—	1559..1747	Fwd
DNA Sequence GCGAATTTCTTATGATTTATGATTTTTATTATTAAATAAGTTATAAAAAAAATAAGTGTATACAAATTTTAAAGTGACTCTTAGGTTTTAAAACGAAAATTCTTATTCTTGAGTAACTCTTTCCTGTAGGTCAGGTTGCTTTCTCAGGTATAGTATGAGGTCGCTCTTATTGACCACACCTCTACCGG