Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | DH6-2 | Length | 4282 bp |
| Type | Cloning vector | Source | Huang DC, Holtz WJ, Maharbiz MM. |
DH6-2 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 ℃.
DH6-2 vector Sequence
LOCUS Exported 4282 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector DH6-2, complete sequence.
ACCESSION JX155232
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4282)
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 4282)
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 4282)
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..4282
/organism="Cloning vector DH6-2"
/mol_type="other DNA"
/db_xref="taxon:1209495"
CDS complement(49..1131)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS complement(49..1008)
/codon_start=1
/product="LacI"
/label=LacI
/protein_id="AFN88249.1"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
promoter complement(1132..1209)
/gene="lacI"
/label=lacI promoter
regulatory 1500..1553
/regulatory_class="promoter"
/note="pLlacO-1"
protein_bind 1502..1518
/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)."
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
gene 1604..2401
/gene="CI"
/label=CI
/note="with mf degradation tag"
CDS 1604..2398
/codon_start=1
/gene="CI"
/product="CI with mf ssrA tag"
/label=CI
/protein_id="AFN88250.1"
/translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
VGKVIASQWPEETFGAANKNEENTNEVPTFMLNAGQANYAFA"
CDS 1604..2314
/codon_start=1
/gene="cI"
/product="phage lambda repressor"
/label=-lambda repressor
/translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
VGKVIASQWPEETFG"
misc_feature 2315..2395
/gene="CI"
/label=mf lon degrdation tag
/note="mf lon degrdation tag"
regulatory 2426..2554
/regulatory_class="terminator"
terminator 2434..2505
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2521..2548
/label=T7Te terminator
/note="phage T7 early transcription terminator"
rep_origin complement(2688..3370)
/direction=LEFT
/label=colE1
/note="colE1"
rep_origin complement(2706..3294)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
regulatory 3376..3481
/regulatory_class="terminator"
terminator 3382..3476
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3497..4156)
/codon_start=1
/product="Cm Resistance"
/label=Cm Resistance
/protein_id="AFN88251.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(4157..4259)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
This page is informational only.