Basic Vector Information
Basic Vector Information | |||
---|---|---|---|
Vector Name | DH7-19 | Antibiotic Resistance | Tetracycline |
Length | 7582 bp | Type | Cloning vector |
Source | Huang DC, Holtz WJ, Maharbiz MM. |
DH7-19 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 ℃.
DH7-19 vector Sequence
LOCUS Exported 7582 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector DH7-19, complete sequence. ACCESSION JX155248 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) AUTHORS . TITLE Direct Submission 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 /organism="Cloning vector DH7-19" /mol_type="other DNA" /db_xref="taxon:1209511" 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 degradation tag" /label=mf-lon with LVA /protein_id="AFN88289.1" /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) regulatory complement(2435..2488) /regulatory_class="promoter" /note="pLtetO-1" 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 2470..2488 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" regulatory 2495..2576 /regulatory_class="promoter" /note="pRM" 5'UTR 2583..2617 gene 2618..3415 /gene="cI-mf-ssra tag" /label=cI-mf-ssra tag CDS 2618..3412 /codon_start=1 /gene="cI-mf-ssra tag" /product="CI with mf Lon degradation tag" /label=cI-mf-ssra tag /protein_id="AFN88290.1" /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV VGKVIASQWPEETFGAANKNEENTNEVPTFMLNAGQANYAFA" CDS 2618..3328 /codon_start=1 /gene="cI" /product="phage lambda repressor" /label=-lambda repressor /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV VGKVIASQWPEETFG" 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 gene 3463..4122 /gene="TetR ssra tag" /label=TetR ssra tag CDS 3463..4119 /codon_start=1 /gene="TetR ssra tag" /product="TetR with LVA degradation tag" /label=TetR ssra tag /protein_id="AFN88291.1" /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAANDENYALVA" CDS 3463..4080 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG" misc_feature 4084..4116 /gene="TetR ssra tag" /label=degradation tag (LVA) /note="degradation tag (LVA)" CDS 4084..4116 /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" regulatory 4147..4275 /regulatory_class="terminator" 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" rep_origin 4277..6505 /label=pSC101 /note="pSC101" CDS complement(4800..5750) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" regulatory 6506..6611 /regulatory_class="terminator" terminator 6512..6606 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(6637..7431) /codon_start=1 /product="Neo Resistance (Kan)" /label=Neo Resistance /protein_id="AFN88292.1" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
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