Basic Vector Information
DH6-1 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 ℃.
DH6-1 vector Sequence
LOCUS 40924_605 4234 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector DH6-1, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4234) AUTHORS Huang DC, Holtz WJ, Maharbiz MM. TITLE A genetic bistable switch utilizing nonlinear protein degradation JOURNAL J Biol Eng 6 (1), 9 (2012) PUBMED 22776405 REFERENCE 2 (bases 1 to 4234) AUTHORS Huang DC, Holtz WJ, Maharbiz MM. TITLE Direct Submission JOURNAL Submitted (10-JUN-2012) Electrical Engineering and Computer Sciences, University of California, Berkeley, 656 Sutardja Dai Hall, Berkeley, CA 94720, USA REFERENCE 3 (bases 1 to 4234) TITLE Direct Submission REFERENCE 4 (bases 1 to 4234) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Eng"; date: "2012"; volume: "6"; issue: "1"; pages: "9" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (10-JUN-2012) Electrical Engineering and Computer Sciences, University of California, Berkeley, 656 Sutardja Dai Hall, Berkeley, CA 94720, USA" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Assembly Method :: GENtle v. 1.9.4 Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..4234 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind complement(15..36) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(52..1131) /label=lacI /note="lac repressor" promoter complement(1132..1209) /label=lacI promoter protein_bind 1502..1518 /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)." protein_bind 1525..1541 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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)." 5'UTR 1569..1603 CDS 1604..2314 /label=lambda repressor /note="phage lambda repressor" CDS 2315..2347 /label=ssrA tag (LVA) /note="C-terminal peptide that mediates degradation in bacteria through the ClpXP and ClpAP proteases (McGinness et al., 2006)" terminator 2386..2457 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2473..2500 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(2658..3246) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator complement(3334..3428) /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3452..4108) /label=CmR /note="chloramphenicol acetyltransferase" promoter complement(4109..4211) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase"
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