Basic Vector Information
- Vector Name:
- pMU1817
- Length:
- 9876 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Holwerda EK, Olson DG, Ruppertsburger NM, Stevenson D
- Promoter:
- URA3
pMU1817 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 ℃.
pMU1817 vector Sequence
LOCUS V016083 9876 bp DNA circular SYN 28-JAN-2019
DEFINITION Exported.
ACCESSION V016083
VERSION V016083
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9876)
AUTHORS Holwerda EK, Olson DG, Ruppertsburger NM, Stevenson D, Murphy SJL.,
Maloney MI, Lanahan A, Amador- Noguez D, Lynd LR.
TITLE Evolutionary response of Clostridium thermocellum to genetic
modifications aimed at improving ethanol production
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9876)
AUTHORS Olson DG, Argyros DA.
TITLE Direct Submission
JOURNAL Submitted (09-OCT-2018) Thayer School of Engineering, Dartmouth
College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE 3 (bases 1 to 9876)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-OCT-2018) Thayer School of Engineering, Dartmouth College, 14
Engineering Drive, Hanover, NH 03755, USA"
FEATURES Location/Qualifiers
source 1..9876
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(294..1094)
/label="URA3"
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
promoter complement(1095..1315)
/label="URA3 promoter"
misc_feature 1343..1846
/label="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
misc_feature complement(1888..2922)
/label="3' homology region"
/note="3' homology region"
misc_feature complement(2923..3765)
/label="5' homology region"
/note="5' homology region"
gene complement(3884..4429)
/gene="hpt"
/label="hpt"
CDS complement(3884..4429)
/codon_start=1
/transl_table=11
/gene="hpt"
/product="hypoxanthine phosphoribosyltransferase"
/label="hpt"
/protein_id="QAT79155.1"
/translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
EKYRNLPFIGVLKPEMYS"
CDS complement(4455..5102)
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
regulatory complement(5103..5630)
/label="gapDH promoter from Clostridium thermocellum"
/note="gapDH promoter from Clostridium thermocellum"
/regulatory_class="promoter"
misc_feature complement(5631..6262)
/label="internal homology region"
/note="internal homology region"
gene complement(6263..6841)
/gene="tdk"
/label="tdk"
CDS complement(6263..6841)
/codon_start=1
/transl_table=11
/gene="tdk"
/product="thymidine dikinase"
/label="tdk"
/protein_id="QAT79157.1"
/translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
ANYDDPIIMVGAKESYEARCRKCHEVPRT"
regulatory complement(6842..7462)
/label="Clostridium thermocellum cbp promoter"
/note="Clostridium thermocellum cbp promoter"
/regulatory_class="promoter"
CDS complement(7572..8573)
/label="repB"
/note="RepB replication protein"
rep_origin complement(9265..9853)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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