Basic Vector Information
mini-CTX2 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 ℃.
mini-CTX2 vector Sequence
LOCUS 40924_1979 6755 bp DNA circular SYN 17-DEC-2018
DEFINITION Integration vector mini-CTX2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6755)
AUTHORS Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
TITLE A site-specific gene integration system for Pseudomonas aeruginosa
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6755)
AUTHORS Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (06-APR-1999) Microbiology, Colorado State University,
Fort Collins, CO 80523, USA
REFERENCE 3 (bases 1 to 6755)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6755)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-APR-1999) Microbiology, Colorado State University, Fort Collins,
CO 80523, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6755
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 827..845
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 858..965
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(974..992)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind complement(1166..1213)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
oriT complement(1420..1528)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
protein_bind complement(2877..2893)
/label=lac operator
/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)."
promoter complement(2901..2930)
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS complement(3174..4253)
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(4254..4331)
/label=lacI promoter
misc_feature 4517..4657
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4843..5431)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5511..6698)
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
promoter complement(join(6746..6755,1..19))
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
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