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mini-CTX2 vector (V009848#)

Basic Vector Information

Basic Vector Information
Vector Name mini-CTX2 Antibiotic Resistance Tetracycline
Length 6755 bp Type Integration vector
Source Hoang TT, Kutchma AJ, Becher A, Schweizer HP.

mini-CTX2 vector Vector Map

mini-CTX26755 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600Flp recombinase binding sitephage CTX attachment site, core sequenceT4 transcription termination signalT3 promotermultiple cloning siteT7 promoterT4 transcription termination signalFlp recombinase binding siteinttrc promoter region with ribosome binding sitelacIlacI promoterbomoritet

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6755 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integration vector mini-CTX2, complete sequence.
ACCESSION   AF140577
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6755
                     /organism="Integration vector mini-CTX2"
                     /lab_host="Pseudomonas aeruginosa"
                     /mol_type="other DNA"
                     /db_xref="taxon:106407"
     protein_bind    238..285
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
                     /note="Flp recombinase target site"
     misc_feature    495..521
                     /label=phage CTX attachment site, core sequence
                     /note="phage CTX attachment site, core sequence"
     regulatory      745..769
                     /regulatory_class="terminator"
                     /note="T4 transcription termination signal"
     regulatory      827..846
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    858..965
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     882..898
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(932..948)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      973..992
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        complement(974..992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1051..1075
                     /regulatory_class="terminator"
                     /note="T4 transcription termination signal"
     protein_bind    1166..1213
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
                     /note="Flp recombinase target site"
     oriT            1413..1672
     oriT            1420..1528
                     /note="incP origin of transfer"
     CDS             complement(1687..2856)
                     /codon_start=1
                     /gene="int"
                     /product="integrase"
                     /label=int
                     /protein_id="AAF06114.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(1687..2856)
                     /gene="int"
                     /label=int
                     /note="modified phage CTX integrase gene"
     regulatory      2860..2946
                     /regulatory_class="promoter"
                     /note="trc promoter region with ribosome binding site"
     protein_bind    2877..2893
                     /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)."
     promoter        complement(2901..2930)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(3171..4253)
                     /codon_start=1
                     /gene="lacI"
                     /product="Lac repressor"
                     /label=lacI
                     /protein_id="AAF06115.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3171..4253)
                     /gene="lacI"
                     /label=lacI
                     /note="GTG start"
     promoter        complement(4254..4331)
                     /gene="lacI"
                     /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(5508..6698)
                     /codon_start=1
                     /gene="tet"
                     /product="Tet protein"
                     /label=tet
                     /protein_id="AAF06116.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(5508..6698)
                     /gene="tet"
                     /label=tet
                     /note="modified tet gene"
     promoter        complement(6746..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"

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