Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | DH6-56 | Length | 5883 bp |
| Type | Cloning vector | Source | Huang DC, Holtz WJ, Maharbiz MM. |
DH6-56 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 ℃.
DH6-56 vector Sequence
LOCUS Exported 5883 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector DH6-56, complete sequence.
ACCESSION JX155234
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5883)
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 5883)
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 5883)
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..5883
/organism="Cloning vector DH6-56"
/mol_type="other DNA"
/db_xref="taxon:1209497"
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="AFN88255.1"
/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..1602
gene 1603..4002
/gene="mfLon"
/label=mfLon
CDS 1603..3999
/codon_start=1
/gene="mfLon"
/product="mf-Lon with ssrA degrdation tag LVA"
/label=mfLon
/protein_id="AFN88256.1"
/translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTKAANDENYALVA"
misc_feature 3964..3996
/gene="mfLon"
/label=degradation tag
/note="degradation tag"
CDS 3964..3996
/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 4027..4155
/regulatory_class="terminator"
terminator 4035..4106
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4122..4149
/label=T7Te terminator
/note="phage T7 early transcription terminator"
rep_origin complement(4289..4971)
/direction=LEFT
/label=colE1
/note="colE1"
rep_origin complement(4307..4895)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
regulatory 4977..5082
/regulatory_class="terminator"
terminator 4983..5077
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(5098..5757)
/codon_start=1
/product="Cm Resistance"
/label=Cm Resistance
/protein_id="AFN88257.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(5758..5860)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
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