Basic Vector Information
- Vector Name:
- PSM2
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6504 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Rahnama M, Forester N, Ariyawansa S, Voisey CR, Johnson LJ, Johnson RD, Fleetwood DJ.
PSM2 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 ℃.
PSM2 vector Sequence
LOCUS 40924_40707 6504 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PSM2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6504)
AUTHORS Rahnama M, Forester N, Ariyawansa S, Voisey CR, Johnson LJ, Johnson
RD, Fleetwood DJ.
TITLE Efficient targeted mutagenesis in Epichloe festucae using a split
marker system
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6504)
AUTHORS Rahnama M, Fleetwood D.
TITLE Direct Submission
JOURNAL Submitted (27-SEP-2016) School of Biological Sciences, University of
Auckland, 3A Symonds Street, Auckland, Auckland 1010, New Zealand
REFERENCE 3 (bases 1 to 6504)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6504)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-SEP-2016) School of Biological Sciences, University of Auckland,
3A Symonds Street, Auckland, Auckland 1010, New Zealand"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6504
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS 513..1454
/codon_start=1
/gene="hph"
/product="hygromycin B phosphotransferase"
/label=hph
/protein_id="APD72153.1"
/translation="SEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALP
IPEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSG
FGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEV
RHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFE
RRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQI
ARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
gene 513..1454
/gene="hph"
/label=hph
terminator 1472..2240
/label=trpC terminator
/note="transcription terminator from the Aspergillus
nidulans trpC gene (Punt et al., 1987)"
primer_bind 2279..2295
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2312..2543
/label=attP1
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
CDS complement(2942..3244)
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(3592..4248)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(4249..4351)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(4496..4727)
/label=attP2
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
promoter complement(4746..4764)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4769..4785)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 4898..5704
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 5854..6442
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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