Basic Vector Information
- Vector Name:
- pEMY128AD_pY128
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4928 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Young EM, Zhao Z, Gielesen BE, Wu L, Benjamin Gordon D, Roubos JA, Voigt CA.
- Promoter:
- TEF
pEMY128AD_pY128 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 ℃.
pEMY128AD_pY128 vector Sequence
LOCUS 40924_17394 4928 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEMY128AD_pY128, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4928)
AUTHORS Young EM, Zhao Z, Gielesen BE, Wu L, Benjamin Gordon D, Roubos JA,
Voigt CA.
TITLE Iterative algorithm-guided design of massive strain libraries,
applied to itaconic acid production in yeast
JOURNAL Metab. Eng. 48, 33-43 (2018)
PUBMED 29753070
REFERENCE 2 (bases 1 to 4928)
AUTHORS Young E, Zhao Z, Gielesen B, Wu L, Gordon DB, Roubos J, Voigt C.
TITLE Direct Submission
JOURNAL Submitted (21-MAY-2018) Genetics, DSM Biotechnology Center,
Alexander Fleminglaan 1, Delft, South Holland 2613AX, Netherlands
REFERENCE 3 (bases 1 to 4928)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4928)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
48, 33-43 (2018)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-MAY-2018) Genetics, DSM Biotechnology Center, Alexander
Fleminglaan 1, Delft, South Holland 2613AX, Netherlands"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4928
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 229..245
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 246..302
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(315..331)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(339..355)
/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)."
promoter complement(363..393)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(408..429)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(659..1247)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
rep_origin 1346..2505
/label=2u ori
/note="yeast 2u plasmid origin of replication"
gene complement(2547..3903)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
CDS complement(4075..4731)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(4732..4835)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
terminator complement(4894..4923)
/label=T3Te terminator
/note="phage T3 early transcription terminator"
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