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psiLentGene Hygromycin vector (V003185#)

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

psiLentGene Hygromycin4601 bp600120018002400300036004200vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)SP6 promoterM13 revlac operatorlac promoterCAP binding siteSV40 promoterHygRsynthetic polyadenylation signaloriAmpRAmpR promoterf1 oriM13 fwd

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

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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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