Basic Vector Information
- Vector Name:
- pNML54
- Antibiotic Resistance:
- Streptomycin
- Length:
- 4147 bp
- Type:
- Binary vector
- Replication origin:
- oriV
- Source/Author:
- Rozwadowski K, Yang W, Kagale S.
pNML54 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 ℃.
pNML54 vector Sequence
LOCUS 40924_33407 4147 bp DNA circular SYN 18-DEC-2018
DEFINITION Binary vector pNML54, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4147)
AUTHORS Rozwadowski K, Yang W, Kagale S.
TITLE Homologous recombination-mediated cloning and manipulation of
genomic DNA regions using Gateway and recombineering systems
JOURNAL BMC Biotechnol. 8, 88 (2008)
PUBMED 19014699
REFERENCE 2 (bases 1 to 4147)
AUTHORS Rozwadowski KL, Yang W, Kagale S.
TITLE Direct Submission
JOURNAL Submitted (21-OCT-2008) Molecular Genetics, Agriculture and
Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan S7N
0X2, Canada
REFERENCE 3 (bases 1 to 4147)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4147)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC
Biotechnol. 8, 88 (2008)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-OCT-2008) Molecular Genetics, Agriculture and Agri-Food Canada,
107 Science Place, Saskatoon, Saskatchewan S7N 0X2, Canada"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4147
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(106..130)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
misc_feature 171..278
/label=MCS
/note="pBluescript multiple cloning site"
misc_feature complement(342..366)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
rep_origin 444..1075
/label=oriV
/note="incP origin of replication"
CDS 1918..2592
/codon_start=1
/gene="aad9"
/product="Aad9"
/label=aad9
/note="spectinomycin resistance"
/protein_id="ACL35309.1"
/translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
KL"
gene 1918..2592
/gene="aad9"
/label=aad9
CDS 2953..4098
/codon_start=1
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
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