Basic Vector Information
pTcGFPN 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 ℃.
pTcGFPN vector Sequence
LOCUS 40924_42759 7584 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pTcGFPN, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7584) AUTHORS Batista M, Marchini FK, Celedon PA, Fragoso SP, Probst CM, Preti H, Ozaki LS, Buck GA, Goldenberg S, Krieger MA. TITLE A high-throughput cloning system for reverse genetics in Trypanosoma cruzi JOURNAL BMC Microbiol. 10 (1), 259 (2010) PUBMED 20942965 REFERENCE 2 (bases 1 to 7584) AUTHORS Batista M, Marchini FK, Celedon PAF., Fragoso SP, Probst CM, Preti H, Ozaki LS, Buck GA, Goldenberg S, Krieger MA. TITLE Direct Submission JOURNAL Submitted (28-APR-2010) Instituto Carlos Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil REFERENCE 3 (bases 1 to 7584) AUTHORS Batista M, Marchini FK, Celedon PAF., Fragoso SP, Probst CM, Preti H, Ozaki LS, Buck GA, Goldenberg S, Krieger MA. TITLE Direct Submission JOURNAL Submitted (18-NOV-2010) Instituto Carlos Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil REFERENCE 4 (bases 1 to 7584) TITLE Direct Submission REFERENCE 5 (bases 1 to 7584) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC Microbiol."; date: "2010"; volume: "10"; issue: "1"; pages: "259" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (28-APR-2010) Instituto Carlos Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (18-NOV-2010) Instituto Carlos Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil" COMMENT SGRef: number: 4; type: "Journal Article" COMMENT On Nov 18, 2010 this sequence version replaced HM162843.1. FEATURES Location/Qualifiers source 1..7584 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(3..458) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 659..1275 /note="derived from Trypanosoma cruzi 18S ribosomal RNA gene promoter region" /regulatory_class="promoter" CDS 1569..2282 /label=EGFP /note="enhanced GFP" protein_bind 2292..2411 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 2441..2471 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 2525..3181 /label=CmR /note="chloramphenicol acetyltransferase" CDS 3526..3828 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" protein_bind complement(3872..3996) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" CDS 4296..5084 /label=NeoR/KanR /note="aminoglycoside phosphotransferase from Tn5" promoter complement(5396..5414) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5435..5451) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5459..5475) /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)." promoter complement(5483..5513) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5528..5549) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5837..6425) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6599..7456) /label=AmpR /note="beta-lactamase" promoter complement(7457..7561) /label=AmpR promoter
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