1136- Desmoplakin-GFP vector (V007420)

Basic Vector Information

      • Vector Name:
      • 1136- Desmoplakin-GFP
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 13661 bp
      • Type:
      • Mammalian Expression
      • Copy Number:
      • High Copy
      • Promoter:
      • CMV
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • CMV-Fwd
      • 3' Primer:
      • EGFP-N

1136- Desmoplakin-GFP vector Vector Map

1136- Desmoplakin-GFP13661 bp600120018002400300036004200480054006000660072007800840090009600102001080011400120001260013200CMV enhancerCMV promoterKozak sequenceSV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRTK-pA-RHSV TK poly(A) signalori

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 ℃.

1136- Desmoplakin-GFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               13661 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    1136- Desmoplakin-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13661)
  AUTHORS   Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, 
            Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew 
            TL, Green KJ
  TITLE     Desmoplakin assembly dynamics in four dimensions: multiple phases 
            differentially regulated by intermediate filaments and actin.
  JOURNAL   J Cell Biol. 2005 Dec 19;171(6):1045-59.
  PUBMED    16365169
REFERENCE   2  (bases 1 to 13661)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2005-12-19"; volume: "171(6)"; pages: "1045-59"
FEATURES             Location/Qualifiers
     source          1..13661
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      9608..9617
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             9614..10333
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             9614..10330
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9659..9680)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9920..9939)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10267..10288
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    10456..10577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10493..10512)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10547..10566
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(10584..11039)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10721..10742)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10933..10952
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        11066..11170
                     /gene="bla"
                     /label=AmpR promoter
     promoter        11172..11529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(11195..11215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11380..11515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11442..11461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             11564..12358
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(11618..11637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     12228..12247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(12546..12565)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    12590..12637
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      12966..13554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13455..13474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"

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