Basic Vector Information
pTcPTPN-NH 1.1 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 ℃.
pTcPTPN-NH 1.1 vector Sequence
LOCUS 40924_42814 7359 bp DNA circular SYN 18-DEC-2018 DEFINITION Protein fusion vector pTcPTPN-NH 1.1, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7359) AUTHORS Kugeratski FG, Batista M, Inoue AH, Ramos BD, Krieger MA, Marchini/ FK. TITLE pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation JOURNAL Mem. Inst. Oswaldo Cruz (2015) In press PUBMED 26200713 REFERENCE 2 (bases 1 to 7359) AUTHORS Kugeratski FG, Batista M, Inoue AH, Ramos BD, Krieger MA, Marchini FK. TITLE Direct Submission JOURNAL Submitted (22-APR-2015) Mass Spectrometry Platform, Instituto Carlos Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil REFERENCE 3 (bases 1 to 7359) TITLE Direct Submission REFERENCE 4 (bases 1 to 7359) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst. Oswaldo Cruz (2015) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (22-APR-2015) Mass Spectrometry Platform, Instituto Carlos Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7359 /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 1599..1772 /label=ProtA /note="IgG-binding unit of Staphylococcus aureus protein A" CDS 1776..1946 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNDAQAPK" CDS 1983..2003 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" protein_bind 2055..2174 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 2204..2234 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 2288..2944 /label=CmR /note="chloramphenicol acetyltransferase" CDS 3289..3591 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" protein_bind complement(3635..3759) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" CDS 4071..4859 /label=NeoR/KanR /note="aminoglycoside phosphotransferase from Tn5" promoter complement(5171..5189) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5210..5226) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5234..5250) /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(5258..5288) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5303..5324) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5612..6200) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6374..7231) /label=AmpR /note="beta-lactamase" promoter complement(7232..7336) /label=AmpR promoter
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