HygR-5' EGFP reporter vector (V009907)

Basic Vector Information

      • Vector Name:
      • HygR-5' EGFP reporter
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6083 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y, Konishi H.

HygR-5' EGFP reporter vector Vector Map

HygR-5' EGFP reporter6083 bp30060090012001500180021002400270030003300360039004200450048005100540057006000CMV enhancerCMV promoterT7 promotervertebrate consensus sequence for strong initiation of translation (Kozak, 1987)SV40 poly(A) signalpartial SV40 largeT; an internal portion of SV40 largeT exon 2M13 revlac operatorlac promoterCAP binding siteoriampRAmpR promoter

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 ℃.

HygR-5' EGFP reporter vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6083 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector HygR-5' EGFP reporter, complete sequence.
ACCESSION   JQ394982
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  AUTHORS   Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y, 
            Konishi H.
  TITLE     A system for the measurement of gene targeting efficiency in human 
            cell lines using an antibiotic resistance-GFP fusion gene
  JOURNAL   BioTechniques 53 (3), 141-152 (2012)
  PUBMED    22963476
REFERENCE   2  (bases 1 to 6083)
  AUTHORS   Konishi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JAN-2012) Aichi Medical University School of Medicine,
            1-1 Yazako-Karimta, Nagakute, Aichi 480-1195, Japan
REFERENCE   3  (bases 1 to 6083)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6083
                     /organism="Cloning vector HygR-5' EGFP reporter"
                     /mol_type="other DNA"
                     /db_xref="taxon:1249021"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      309..810
                     /regulatory_class="promoter"
                     /note="Human cytomegalovirus (CMV) major immediate early 
                     promoter"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      936..945
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             942..2524
                     /codon_start=1
                     /gene="HygR-5' EGFP"
                     /product="HygR-5' EGFP"
                     /label=HygR-5' EGFP
                     /note="fusion protein of hygromycin B phosphotransferase 
                     and 3'-truncated enhanced GFP"
                     /protein_id="AFV73966.1"
                     /translation="MDRSGKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVG
                     GRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQ
                     DLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSE
                     AMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSL
                     VDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRP
                     RAKEPPPPPDRSYVSGFEFATMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA
                     TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQER
                     TIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMAD
                     KQKNGIKVNF"
     gene            942..2524
                     /gene="HygR-5' EGFP"
                     /label=HygR-5' EGFP
     CDS             957..1973
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     regulatory      2019..2028
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    2673..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2704..2754
                     /label=SV40 early mRNA polyA
                     /note="SV40 early mRNA polyA"
     misc_feature    2865..3883
                     /note="partial SV40 largeT; an internal portion of SV40 
                     largeT exon 2"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3950..3966
                     /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(3974..4004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4019..4040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4328..4916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5087..5947)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicilin resistance protein"
                     /protein_id="AFV73967.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5087..5947)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(5948..6052)
                     /gene="bla"
                     /label=AmpR promoter

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