Cip-pAct1-Mnase

162556 Synthetic Yeast Expression

Plasmid Information

Purpose
NheI-MluI-digested 3xFLAG-MNase-SV40 fragment was cloned into the CIp-pACT1-CYC vector to ensure constitutive expression of MNase in C. albicans.
Depositor
Adnane Sellam
Article
Tebbji et al mSphere. 2020 Oct 14;5(5). pii: 5/5/e006
Insert
3FLAG-Mnase-SV40
Promoter
Actine
Available Since
Jan. 29, 2021
Availability
Available

Sequence Information

Length
6,804 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
3xFLAG — three tandem FLAG(R) epitope tags, foll… 1715..1781 Fwd
DNA Sequence
GACTACAAAGACCACGACGGAGACTATAAGGATCACGATATTGACTATAAGGACGATGATGATAAA
Protein Sequence
DYKDHDGDYKDHDIDYKDDDDK
MNase — micrococcal nuclease from Staphylococcu… 1787..2234 Fwd
DNA Sequence
GCCACCTCAACAAAAAAACTTCACAAAGAACCTGCAACCTTAATAAAGGCAATTGACGGGGACACTGTCAAATTGATGTATAAAGGTCAGCCAATGACCTTCCGTTTACTTTTAGTCGATACTCCTGAGACTAAGCATCCTAAGAAAGGTGTAGAGAAATATGGTCCTGAAGCCAGTGCTTTCACAAAGAAAATGGTTGAGAATGCAAAGAAAATTGAGGTAGAGTTCGACAAAGGACAAAGAACCGACAAGTATGGTAGAGGGTTGGCCTACATCTACGCAGATGGAAAGATGGTTAACGAAGCATTAGTCCGACAGGGTTTGGCTAAAGTTGCCTATGTGTATAAGCCTAACAATACCCACGAACAGCACCTTCGTAAGTCAGAAGCTCAAGCTAAAAAAGAAAAGTTGAATATCTGGTCAGAGGATAATGCAGACTCTGGTCAA
Protein Sequence
ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETKHPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALVRQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ
SV40 NLS — nuclear localization signal of SV40 (si… 2237..2258 Fwd
DNA Sequence
CCTAAGAAAAAACGTAAAGTA
Protein Sequence
PKKKRKV
AmpR bla beta-lactamase 5395..6256 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 4
Label Gene Product Position Strand
lac promoter — — 534..565 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
T3 promoter — — 633..652 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
T7 promoter — — 4626..4645 Rev
DNA Sequence
TAATACGACTCACTATAGG
AmpR promoter bla — 5290..5395 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
CAP binding site — — 498..520 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator — — 572..589 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
rep_origin 2
Label Gene Product Position Strand
ori — — 0..6804 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
f1 ori — — 4808..5264 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT