Basic Vector Information
- Vector Name:
- pMB1_MmPylRS-AfTyrRS.p-I-Phe
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5594 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Robertson WE, Funke LFH., de la Torre D, Fredens J
pMB1_MmPylRS-AfTyrRS.p-I-Phe 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 ℃.
pMB1_MmPylRS-AfTyrRS.p-I-Phe vector Sequence
LOCUS 62056_16550 5594 bp DNA circular SYN 12-JUN-2021
DEFINITION Cloning vector pMB1_MmPylRS/AfTyrRS.p-I-Phe, complete sequence.
ACCESSION MW879727
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5594)
AUTHORS Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS,
Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
S, Salmond GPC., Chin JW.
TITLE Sense Codon Reassignment Enables Viral Resistance and Encoded
Polymer Synthesis
JOURNAL Science (2021) In press
REFERENCE 2 (bases 1 to 5594)
AUTHORS Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS,
Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
S, Salmond GPC., Chin JW.
TITLE Direct Submission
JOURNAL Submitted (06-APR-2021) Protein and Nucleic Acid Chemistry, MRC
Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge,
Cambridgeshire CB2 0QH, United Kingdom
REFERENCE 3 (bases 1 to 5594)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science
(2021) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-APR-2021) Protein and Nucleic Acid Chemistry, MRC Laboratory of
Molecular Biology, Francis Crick Avenue, Cambridge, Cambridgeshire
CB2 0QH, United Kingdom"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5594
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 197..241
/label=S-Tag
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
terminator 293..340
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(514..1326)
/label=KanR
/note="aminoglycoside phosphotransferase"
promoter complement(1327..1418)
/label=AmpR promoter
rep_origin complement(1482..2071)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
gene 2155..2180
/gene="glnS"
/label=glnS
regulatory 2155..2180
/gene="glnS"
/label=GlnS promoter
/note="GlnS promoter"
/regulatory_class="promoter"
CDS 2225..3586
/gene="pylS"
/label=pylS
/note="Pyrrolysine--tRNA ligase from Methanosarcina mazei
(strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 /
OCM 88). Accession#: Q8PWY1"
CDS 3639..4607
/gene="tyrS"
/label=tyrS
/note="Tyrosine--tRNA ligase from Archaeoglobus fulgidus
(strain ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 /
VC-16). Accession#: O29482"
regulatory 5143..5193
/label=lpp promoter
/note="lpp promoter"
/regulatory_class="promoter"
tRNA 5200..5271
/product="tRNA-Ser"
/label=pylToptserU(CGA)
/note="pylToptserU(CGA)"
tRNA 5311..5385
/product="tRNA-Tyr"
/label=Af tRNA TyrA01(CUA)
/note="Af tRNA TyrA01(CUA)"
gene 5392..5420
/gene="rrnC"
/label=rrnC
regulatory 5392..5420
/gene="rrnC"
/label=rrnC term
/note="rrnC term"
/regulatory_class="terminator"
primer_bind complement(5430..5446)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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