Basic Vector Information
- Vector Name:
- hygII-pSmRYNE(R)
- Antibiotic Resistance:
- Kanamycin
- Length:
- 13281 bp
- Type:
- BiFC expression vector
- Replication origin:
- oriV
- Source/Author:
- Offenborn JN, Waadt R, Kudla J.
- Promoter:
- MAS
hygII-pSmRYNE(R) 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 ℃.
hygII-pSmRYNE(R) vector Sequence
LOCUS 40924_1369 13281 bp DNA circular SYN 17-DEC-2018 DEFINITION BiFC expression vector hygII-pSmRYNE(R), complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13281) AUTHORS Offenborn JN, Waadt R, Kudla J. TITLE Visualization and translocation of ternary Calcineurin-A/Calcineurin-B/Calmodulin-2 protein complexes by dual-color trimolecular fluorescence complementation JOURNAL New Phytol. (2015) In press PUBMED 25919910 REFERENCE 2 (bases 1 to 13281) AUTHORS Offenborn JN, Waadt R, Kudla J. TITLE Direct Submission JOURNAL Submitted (22-MAY-2015) University of Muenster, Institute for Biology and Plant Biotechnology, Schlossplatz 7, Muenster 48149, Germany REFERENCE 3 (bases 1 to 13281) TITLE Direct Submission REFERENCE 4 (bases 1 to 13281) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "New Phytol. (2015) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (22-MAY-2015) University of Muenster, Institute for Biology and Plant Biotechnology, Schlossplatz 7, Muenster 48149, Germany" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..13281 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 6..616 /label=oriV /note="origin of replication for the bacterial F plasmid" mobile_element complement(1317..2084) /label=IS1 /note="prokaryotic transposable element" CDS 2276..3067 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" CDS 3369..4514 /codon_start=1 /label=trfA /note="trans-acting replication protein that binds to and activates oriV" /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" misc_feature 6113..6137 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" primer_bind 6755..6771 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator complement(6784..6984) /label=gene 7 terminator /note="transcription terminator for octopine-type Ti plasmid gene 7" CDS complement(7026..8048) /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGAFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" promoter complement(8102..8281) /label=NOS promoter /note="nopaline synthase promoter" promoter 9422..9802 /label=MAS promoter /note="mannopine synthase promoter (Velten et al., 1984)" rep_origin 10000..10624 /label=oriV /note="incP origin of replication" misc_feature 10804..10828 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 11293..11940 /codon_start=1 /label=TetR /note="tetracycline resistance regulatory protein" /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD" oriT 12624..12733 /label=oriT /note="incP origin of transfer" CDS 12766..13134 /codon_start=1 /label=traJ /note="oriT-recognizing protein" /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP"
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