Basic Vector Information
DH7-23 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 ℃.
DH7-23 vector Sequence
LOCUS 40924_660 7582 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector DH7-23, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7582) 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 7582) 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 7582) TITLE Direct Submission REFERENCE 4 (bases 1 to 7582) 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..7582 /mol_type="other DNA" /organism="synthetic DNA construct" gene complement(1..2400) /gene="mf-lon with LVA" /label=mf-lon with LVA CDS complement(4..2400) /codon_start=1 /gene="mf-lon with LVA" /product="mf-Lon with LVA tag" /label=mf-lon with LVA /protein_id="AFN88305.1" /translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTKAANDENYALVA" misc_feature complement(7..39) /gene="mf-lon with LVA" /label=degradation tag (LVA) /note="degradation tag (LVA)" CDS complement(7..39) /codon_start=1 /product="C-terminal peptide that mediates degradation in bacteria through the ClpXP and ClpAP proteases (McGinness et al., 2006)" /label=ssrA tag (LVA) /note="mutant LVA variant that confers accelerated degradation under some conditions (Andersen et al., 1998)" /translation="AANDENYALVA" 5'UTR complement(2401..2434) protein_bind 2445..2463 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind complement(2470..2488) /label=tet operator /note="bacterial operator O2 for the tetR and tetA genes" regulatory 2495..2576 /label=pRM /note="pRM" /regulatory_class="promoter" 5'UTR 2583..2617 CDS 2618..3328 /label=lambda repressor /note="phage lambda repressor" misc_feature 3329..3409 /gene="cI-mf-ssra tag" /label=mf-Lon ssrA degradation tag /note="mf-Lon ssrA degradation tag" 5'UTR 3422..3462 CDS 3463..4083 /label=TetR /note="tetracycline repressor TetR" CDS 4084..4116 /label=ssrA tag (LVA) /note="C-terminal peptide that mediates degradation in bacteria through the ClpXP and ClpAP proteases (McGinness et al., 2006)" terminator 4155..4226 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4242..4269 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4803..5750) /label=Rep101 /note="RepA protein needed for replication with the pSC101 origin" rep_origin complement(5798..6020) /direction=LEFT /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator complement(6512..6606) /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(6640..7431) /label=NeoR/KanR /note="aminoglycoside phosphotransferase"
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