pTcGFPN vector (V002794)

Basic Vector Information

      • Vector Name:
      • pTcGFPN
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7584 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Batista M, Marchini FK, Celedon PA, Fragoso SP, Probst CM, Preti H, Ozaki LS, Buck GA, Goldenberg S, Krieger MA.

pTcGFPN vector Vector Map

pTcGFPN7584 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500f1 oriM13 fwdT7 promoterderived from Trypanosoma cruzi 18S ribosomal RNA gene promoter regionEGFPattR1lac UV5 promoterCmRccdBattR2NeoR/KanRT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR 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 ℃.

pTcGFPN vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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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