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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