pTcmKateN-NH-2.0 vector (V015636)

LOCUS       62056_21060        8055 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcmKateN-NH 2.0, complete sequence.
ACCESSION   MT180307
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  AUTHORS   Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
  TITLE     Characterising ISWI chromatin remodeler in Trypanosoma cruzi
  JOURNAL   Mem. Inst. Oswaldo Cruz 115, e190457 (2020)
  PUBMED    32428081
REFERENCE   2  (bases 1 to 8055)
  AUTHORS   Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz 
            Mader, Curitiba 81350010, Brasil
REFERENCE   3  (bases 1 to 8055)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst. 
            Oswaldo Cruz 115, e190457 (2020)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz Mader, 
            Curitiba 81350010, Brasil"
FEATURES             Location/Qualifiers
     source          1..8055
                     /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      706..1322
                     /note="Trypanosoma cruzi 18S ribosomal RNA promoter region"
                     /regulatory_class="promoter"
     misc_feature    1335..1839
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511469.50 CDS"
     CDS             1849..2643
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    2653..2970
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 CDS"
     CDS             2983..3678
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     protein_bind    3688..3812
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3837..3867
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3921..4577
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4922..5224
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(5268..5392)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    5416..5836
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     promoter        complement(5867..5885)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5906..5922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5930..5946)
                     /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(5954..5984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5999..6020)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6308..6896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7070..7927)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7928..8032)
                     /label=AmpR promoter