MTS-mCherry-GFP1-10-Hyg-N1 vector (V001370)

Basic Vector Information

      • Vector Name:
      • MTS-mCherry-GFP1-10-Hyg-N1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6655 bp
      • Type:
      • Mammalian Expression
      • Copy Number:
      • High Copy
      • Promoter:
      • CMV
      • Cloning Method:
      • Restriction Enzyme

MTS-mCherry-GFP1-10-Hyg-N1 vector Vector Map

MTS-mCherry-GFP1-10-Hyg-N16655 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600SV40 promoterHygRSV40 poly(A) signalpBRforEcoAmpR promoterAmpRoriCMV enhancerCMV promoterKozak sequenceSV40 poly(A) signalf1 oriAmpR 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 ℃.

MTS-mCherry-GFP1-10-Hyg-N1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6655 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses mCherry-GFP1-10 in mammalian cell mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    MTS-mCherry-GFP1-10-Hyg-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  AUTHORS   Ruan L, Zhou C, Jin E, Kucharavy A, Zhang Y, Wen Z, Florens L, Li R
  TITLE     Cytosolic proteostasis through importing of misfolded proteins into 
            mitochondria.
  JOURNAL   Nature. 2017 Mar 16;543(7645):443-446. doi: 10.1038/nature21695. 
            Epub 2017 Mar 1.
  PUBMED    28241148
REFERENCE   2  (bases 1 to 6655)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature21695"; journalName: "Nature"; date: "2017-03-16- 
            16"; volume: "543"; issue: "7645"; pages: "443-446"
FEATURES             Location/Qualifiers
     source          1..6655
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        69..426
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(92..112)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      277..412
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     339..358
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             434..1459
                     /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    1576..1657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1685..1703)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1771..1875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1876..2736
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2094..2113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2922..3510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3411..3430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3685..3988
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3989..4192
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4143..4163
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4497..4506
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    6008..6129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     6099..6118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6136..6591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6273..6294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6485..6504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6618..6655,1..67)
                     /gene="bla"
                     /label=AmpR promoter

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