STOP-eGFP-ROSA26TV vector (V012739)

Basic Vector Information

      • Vector Name:
      • STOP-eGFP-ROSA26TV
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 14367 bp
      • Source/Author:
      • Klaus Rajewsky
      • Copy Number:
      • High copy number
      • Expression Method:
      • Transient

STOP-eGFP-ROSA26TV vector Vector Map

STOP-eGFP-ROSA26TV14367 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000Rosa26 left armAd SAloxPHSV TK poly(A) signalNeoR/KanRsmall t intronSV40 NLSSV40 poly(A) signalloxPFRTIRESEGFPFRTSK primerbGH poly(A) signalPGK promoterDTAsmall t intronbGH poly(A) signalT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterf1 oriM13 fwdT7 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 ℃.

STOP-eGFP-ROSA26TV vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               14367 bp ds-DNA     circular SYN 11-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    STOP-eGFP-ROSA26TV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14367)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14367
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    5..1086
                     /note="Rosa26 left arm"
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    1093..1196
                     /note="Ad SA"
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     protein_bind    1207..1240
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    1247..1295
                     /note="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(1302..2105)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     intron          2919..2984
                     /note="small t intron"
                     /note="simian virus 40 (SV40) small t antigen intron"
     CDS             3114..3134
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     polyA_signal    3559..3693
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     protein_bind    3713..3746
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    3779..3826
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    3865..4416
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4428..5147
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    5172..5219
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5244..5260)
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5279..5503
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        9897..10387
                     /note="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             10414..10983
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphteria toxin Fragment A"
                     /note="DTA"
                     /translation="DDVVILLIFCMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDD
                     DWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELG
                     LSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEIN
                     FETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          11060..11125
                     /note="small t intron"
                     /note="simian virus 40 (SV40) small t antigen intron"
     polyA_signal    11199..11423
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(11522..11540)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11561..11577)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11585..11601
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(11609..11639)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    11654..11675
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(11963..12551)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12722..13582)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(13583..13687)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(13713..14168)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     14310..14326
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        14336..14354
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"

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