Basic Vector Information
Basic Vector Information | |||
---|---|---|---|
Vector Name | psiLentGene Hygromycin | Antibiotic Resistance | Ampicillin |
Length | 4601 bp | Type | Cloning vector |
Source | Virdugirene J. |
psiLentGene Hygromycin 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 ℃.
psiLentGene Hygromycin vector Sequence
LOCUS Exported 4601 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector psiLentGene Hygromycin, complete sequence. ACCESSION AY508734 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4601) AUTHORS Virdugirene J. TITLE siLentGene U6 Hairpin Cloning Systems Technical Manual #TM247 JOURNAL Unpublished REFERENCE 2 (bases 1 to 4601) AUTHORS Virdugirene J. TITLE Direct Submission JOURNAL Submitted (18-DEC-2003) Scientific Communications, Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711, USA REFERENCE 3 (bases 1 to 4601) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..4601 /organism="Cloning vector psiLentGene Hygromycin" /mol_type="other DNA" /db_xref="taxon:268224" regulatory 33..42 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 37..46 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(140..158) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(176..192) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 200..216 /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(224..254) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 269..290 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 465..822 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 673..808 /label=SV40 ori /note="SV40 origin of replication" CDS 863..1888 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 1923..1971 /note="synthetic polyadenylation signal" rep_origin complement(2164..2752) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2923..3783) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3784..3888) /gene="bla" /label=AmpR promoter rep_origin complement(3966..4421) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(4398..180) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA RTDRPSQQLRSLNGEWTRPVAAH" primer_bind 4562..4578 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 4585..2 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase"
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