pDUAL-pAct1-Sp_IntAct-tADH1

Plasmid Information

Purpose

Low-copy centromeric plasmids which expresses Schizosaccharomyces pombe IntAct (actin internally tagged with ALFA tag between T229 and A230 amino acids)

Depositor

Saravanan Palani

Article

van Zwam et al PLoS Biol. 2024 Mar 11;22(3):e3002551. d

Insert

Sp_IntAct

Sequence Information

Length

10,088 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 3

Label	Gene	Product	Position	Strand
ALFA-tag	—	rationally designed epitope tag that fo…	1914..1953	Fwd
DNA Sequence AGTAGATTAGAAGAAGAACTTCGTAGACGTCTTACTGAA Protein Sequence SRLEEELRRRLTE
S. pombe ura4	Schizosaccharomyces pombe ura4	orotidine 5'-phosphate decarboxylase, r…	4274..5069	Rev
DNA Sequence ATGGATGCTAGAGTATTTCAAAGCTATTCAGCTAGAGCTGAGGGGATGAAAAATCCCATTGCCAAGGAATTGTTGGCTTTGATGGAAGAAAAGCAAAGCAACTTGTCAGTCGCGGTCGATTTGACGAAGAAATCCGAAATCTTAGAATTGGTAGATAAAATTGGACCCTATGTCTGTGTTATCAAGACACATATTGACGTTGTCGAGGATTTCGACCAGGATATGGTAGAAAAACTGGTGGCCTTAGGTAAAAAGCATCGTTTTCTTATCTTTGAGGATCGCAAATTCGCAGACATTGGAAATACCGTCAAGCTACAATATGCATCTGGTGTGTACAAAATTGCTTCTTGGGCTCATATCACAAATTGCCATACAGTGCCAGGCGAGGGTATTATACAAGGCCTCAAAGAAGTTGGTTTACCTTTGGGACGTGGTCTCTTGCTTTTGGCTGAAATGTCTTCCAAAGGCTCTTTGGCTACTGGTTCCTACACAGAGAAAACCTTAGAATGGTTTGAGAAGCATACCGATTTTTGCTTTGGCTTTATAGCTGGTCGTCGATTTCCTAACCTTCAAAGCGACTACATAACTATGTCCCCTGGTATCGGCTTGGATGTTAAAGGAGACGGGCTGGGACAGCAATATCGTACTCCTGAAGAAGTGATTGTAAACTGCGGTAGCGACATCATCATTGTTGGTCGTGGAGTCTATGGAGCTGGTCGTAATCCTGTTGTCGAAGCCAAGAGATATAGAGAAGCTGGTTGGAAGGCATATCAGCAAAGACTTTCTCAGCATTAA Protein Sequence MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKSEILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVKLQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSYTEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIVNCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH
AmpR	bla	beta-lactamase	8254..9115	Fwd
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

promoter 2

Label	Gene	Product	Position	Strand
lac promoter	—	—	109..140	Fwd
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter	bla	—	8149..8254	Fwd
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

protein_bind 2

Label	Gene	Product	Position	Strand
CAP binding site	—	—	73..95	Fwd
DNA Sequence TAATGTGAGTTAGCTCACTCAT
lac operator	—	—	147..164	Fwd
DNA Sequence TTGTGAGCGGATAACAA

rep_origin 3

Label	Gene	Product	Position	Strand
ARS1	—	—	5677..6461	Fwd
DNA Sequence TAGGCATTTTGTTTAGTTAAAGGTACTTAAGTAATTAGTATAAACGAACCAATTTTATAATCAGGAAGTTAAGTGAATGGTAGCACATGTCGTAAAAATTGTGAATTTTTATTGAATAATATTTTAAATACAAGCCTTTCTAGACTAGGTATACTCATAAACATATATGAGCAAAAGGATAGAGGAGATTACATTGCATCTTCTACAAATTTATTTATTGCCCTTTACTGAAAAATTAAATAATGAGTACTAAATGATAAAAAGCGCTCAGTACAAGAAAGATTGCAAAAATATTGCATTCTTCATGAATTAAAGTTGCATATAAGGCATATTGAAAGTAATAGTACTAAAACAGCAGTTAGCGAAAATTAATAGAATTATATTCGCAAGACAATTGTGACATTAAATTAAAAAATTGTAGAATTTTTACTATCCTCTTTAACGCCATGAGCCTTTATAAAAAGGTTAAATTAGTTTTAACATTCTTTTTTTGAGTAAGAGTTTACAATTTATCAAAACCTGTGTTATTATATTCATTAGTTTCAATTTATTAGCATCTAGAGAAAAATCAATTGGCAGTTACATTGTTGGAATTTATGAAGAAAAAGAATCTACAACGGAGAATAAGTTGCTGATCGCTTTCCCTAAAATTGTATATTTTGCTGAGCTTATTTTGACATTTCGTTGAAGTTTTCTCTAATTCGCATTCATTTTAAGTAAAACAATGAGAAATAAAATTACAAAAAATACAAATTAAAATACAATTTTTAGCTATAATTATAGA
f1 ori	—	—	7412..7868	Fwd
DNA Sequence ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTT
ori	—	—	9285..9874	Fwd
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
ADH1 terminator	S. cerevisiae ADH1	—	2423..2611	Fwd
DNA Sequence GCGAATTTCTTATGATTTATGATTTTTATTATTAAATAAGTTATAAAAAAAATAAGTGTATACAAATTTTAAAGTGACTCTTAGGTTTTAAAACGAAAATTCTTATTCTTGAGTAACTCTTTCCTGTAGGTCAGGTTGCTTTCTCAGGTATAGTATGAGGTCGCTCTTATTGACCACACCTCTACCGG