Basic Vector Information
FOXP3 vector Vector Map
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 ℃.
FOXP3 vector Sequence
LOCUS 40924_860 9841 bp DNA circular SYN 17-DEC-2018 DEFINITION Expression vector FOXP3, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9841) AUTHORS Masiuk KE, Laborada J, Grazia Roncarolo M, Hollis RP, Kohn DB. TITLE Hematopoietic Stem Cell Gene Therapy for IPEX Syndrome JOURNAL Unpublished REFERENCE 2 (bases 1 to 9841) AUTHORS Masiuk KE, Laborada J, Grazia Roncarolo M, Hollis RP, Kohn DB. TITLE Direct Submission JOURNAL Submitted (02-OCT-2018) Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, 610 Charles E. Young Dr. East, Los Angeles, CA 90095, USA REFERENCE 3 (bases 1 to 9841) TITLE Direct Submission REFERENCE 4 (bases 1 to 9841) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-OCT-2018) Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, 610 Charles E. Young Dr. East, Los Angeles, CA 90095, USA" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..9841 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 27..131 /label=AmpR promoter CDS 132..989 /label=AmpR /note="beta-lactamase" rep_origin 1163..1751 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2039..2060 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2075..2105 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2113..2129 /label=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)." primer_bind 2137..2153 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2174..2192 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" enhancer 2293..2672 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2673..2876 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 2891..3071 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2891..2986 /label=R region LTR /note="R region LTR" misc_feature 2897..3071 /label=U5 region LTR /note="U5 region LTR" misc_feature 3118..3243 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3736..3969 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 4152..4196 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" CDS 4345..4386 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 4489..4606 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" regulatory 4655..4892 /label=CNS1 /note="CNS1" /regulatory_class="enhancer" regulatory 4893..5252 /label=CNS2 /note="CNS2" /regulatory_class="enhancer" regulatory 5252..5470 /label=CNS3 /note="CNS3" /regulatory_class="enhancer" regulatory 5471..5853 /label=FOXP3 /note="FOXP3" /regulatory_class="promoter" 5'UTR 5854..6041 CDS 6042..7334 /gene="FOXP3" /label=FOXP3 /note="Forkhead box protein P3 from Homo sapiens. Accession#: Q9BZS1" 3'UTR 7338..8213 misc_feature 8291..8331 /label=U3 region LTR (first half) /note="U3 region LTR (first half)" misc_feature 8332..8597 /label=A2 insulator /note="A2 insulator" misc_feature 8598..8614 /label=U3 region LTR (second half) /note="U3 region LTR (second half)" LTR 8615..8795 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 8867..9000 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 9027..9162 /label=SV40 ori /note="SV40 origin of replication" promoter complement(9195..9213) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(9223..9239) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 9384..9839 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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