Basic Vector Information
- Vector Name:
- pNS1MB
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8218 bp
- Type:
- Yeast display vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Uchanski T, Masiulis S, Fischer B, Kalichuk V
- Promoter:
- GAL1,10
pNS1MB vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pNS1MB vector Sequence
LOCUS 62056_18250 8218 bp DNA circular SYN 07-JAN-2021
DEFINITION Yeast display vector pNS1MB, complete sequence.
ACCESSION MT543227
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8218)
AUTHORS Uchanski T, Masiulis S, Fischer B, Kalichuk V, Lopez-Sanchez U,
Zarkadas E, Weckener M, Sente A, Ward P, Wohlkoenig A, Zoegg T,
Remaut H, Naismith JH, Nury H, Vranken W, Aricescu AR, Pardon E,
Steyaert J.
TITLE Megabodies expand the nanobody toolkit for protein structure
determination by single-particle cryo-EM
JOURNAL Nat Methods 18 (1), 60-68 (2021)
REFERENCE 2 (bases 1 to 8218)
AUTHORS Steyaert J, Uchanski T, Pardon E.
TITLE Direct Submission
JOURNAL Submitted (31-MAY-2020) VIB-VUB Center for Structural Biology, VIB,
Pleinlaan 2, Brussels 1050, Belgium
REFERENCE 3 (bases 1 to 8218)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods"; date: "2021"; volume: "18"; issue: "1"; pages: "60-68"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(31-MAY-2020) VIB-VUB Center for Structural Biology, VIB, Pleinlaan
2, Brussels 1050, Belgium"
FEATURES Location/Qualifiers
source 1..8218
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(70..573)
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 610..714
/label=AmpR promoter
CDS 715..1572
/label=AmpR
/note="beta-lactamase"
rep_origin 1746..2334
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2622..2643
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2658..2688
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2696..2712
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 2720..2736
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2757..2775
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2940..3592
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
misc_feature 3632..3898
/label=appS4 leader sequence
/note="appS4 leader sequence"
misc_feature 3899..3937
/label=NS1 monobody beta-strain A (residues 4-16)
/note="NS1 monobody beta-strain A (residues 4-16)"
misc_feature 3938..3940
/label=Phe residue
/note="Phe residue"
misc_feature 3941..5161
/label=cHopQ scaffold protein
/note="cHopQ scaffold protein"
misc_feature 5162..5164
/label=Gly residue
/note="Gly residue"
misc_feature 5165..5401
/label=NS1 monobody C-terminal part (residues 19-97)
/note="NS1 monobody C-terminal part (residues 19-97)"
misc_feature 5408..5509
/label=linker
/note="linker"
misc_feature 5510..5716
/label=Aga2p anchor protein
/note="Aga2p anchor protein"
CDS 5738..5971
/label=ACP
/note="acyl carrier protein (ACP) from E. coli"
misc_feature 5975..6004
/label=Myc-tag
/note="Myc-tag"
regulatory 6018..6324
/label=MFa1
/note="MFa1"
/regulatory_class="terminator"
promoter complement(6339..6357)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6364..6380)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6525..6980
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(7080..7751)
/label=TRP1
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
promoter complement(7752..8032)
/label=TRP1 promoter
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