Basic Vector Information
- Vector Name:
- pET-19b_MMLV-RT.gb
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7789 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- E. coli
- Promoter:
- T7
- 5' Primer:
- T7:TAATACGACTCACTATAGGG
- Growth Temperature:
- 30℃
pET-19b_MMLV-RT.gb 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 ℃.
pET-19b_MMLV-RT.gb vector Sequence
LOCUS 62056_9510 7789 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 7789) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7789 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 204..222 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 223..247 /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 262..284 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 297..323 /codon_start=1 /label=9xHis /note="9xHis affinity tag" /translation="HHHHHHHHH" CDS 342..356 /codon_start=1 /label=enterokinase site /note="enterokinase recognition and cleavage site" /translation="DDDDK" terminator 2507..2554 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(2729..2757) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" promoter 2870..2974 /label=AmpR promoter CDS 2975..3832 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4006..4594 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(4780..4919) /label=bom /note="basis of mobility region from pBR322" CDS complement(5024..5212) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" protein_bind complement(6490..6511) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(6527..7606) /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(7607..7684) /label=lacI promoter
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