Basic Vector Information
- Vector Name:
- D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10037 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- E. coli
- Copy Number:
- Low
- Promoter:
- T7
- Growth Temperature:
- 37℃
D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA 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 ℃.
D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA vector Sequence
LOCUS 62056_616 10037 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10037) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..10037 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 27..45 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 46..70 /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)." RBS 85..107 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 126..143 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 150..1247 /codon_start=1 /label=MBP /note="maltose binding protein from E. coli" /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW YAVRTAVINAASGRQTVDEALKDAQT" protein_bind 1266..1290 /label=attB1 /note="recombination site for the Gateway(R) BP reaction" CDS 1311..1331 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="ENLYFQG" protein_bind complement(4751..4775) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" CDS 4789..4806 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" terminator 4873..4920 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 4957..5412 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5438..5542 /label=AmpR promoter CDS 5543..6400 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6574..7162 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(7348..7487) /label=bom /note="basis of mobility region from pBR322" CDS complement(7592..7780) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" protein_bind complement(8555..8576) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(8592..9671) /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(9672..9749) /label=lacI promoter
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