pIBB103 vector (V005457)

LOCUS       40924_25306        8505 bp DNA     circular SYN 18-DEC-2018
DEFINITION  RNA interference vector pIBB103, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8505)
  AUTHORS   Bose I, Doering TL.
  TITLE     Efficient implementation of RNA interference in the pathogenic yeast
            Cryptococcus neoformans
  JOURNAL   J. Microbiol. Methods 86 (2), 156-159 (2011)
  PUBMED    21554906
REFERENCE   2  (bases 1 to 8505)
  AUTHORS   Bose I, Reilly MC, Skowyra ML, Doering TL.
  TITLE     A convergent promoter vector for RNAi in Cryptococcus neoformans 
            that allows 5-FOA selection for colonies with efficient RNAi
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 8505)
  AUTHORS   Skowyra ML, Doering TL.
  TITLE     RNA interference in Cryptococcus neoformans
  JOURNAL   Unpublished
REFERENCE   4  (bases 1 to 8505)
  AUTHORS   Bose I, Reilly MC, Skowyra ML, Doering TL.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-NOV-2010) Molecular Microbiology, Washington 
            University School of Medicine, 660 S. Euclid Ave. Campus Box 8230, 
            St. Louis, MO 63110, USA
REFERENCE   5  (bases 1 to 8505)
  AUTHORS   Bose I, Reilly MC, Skowyra ML, Doering TL.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2011) Molecular Microbiology, Washington 
            University School of Medicine, 660 S. Euclid Ave. Campus Box 8230, 
            St. Louis, MO 63110, USA
REFERENCE   6  (bases 1 to 8505)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 8505)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2011"; volume: "86"; issue: "2"; pages: 
            "156-159"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "Submitted 
            (01-NOV-2010) Molecular Microbiology, Washington University School 
            of Medicine, 660 S. Euclid Ave. Campus Box 8230, St. Louis, MO 
            63110, USA"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (24-MAY-2011) Molecular Microbiology, Washington University School 
            of Medicine, 660 S. Euclid Ave. Campus Box 8230, St. Louis, MO 
            63110, USA"
COMMENT     SGRef: number: 6; type: "Journal Article"
COMMENT     On May 24, 2011 this sequence version replaced HQ455038.1.
FEATURES             Location/Qualifiers
     source          1..8505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /label=AmpR promoter
     CDS             126..983
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1157..1745
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2069..2099
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2223..2259
                     /note="telomeric repeat; derived from Cryptococcus
                     neoformans var. neoformans JEC21"
     gene            complement(2294..3047)
                     /gene="npt1"
                     /label=npt1
     CDS             complement(2580..2936)
                     /codon_start=1
                     /gene="npt1"
                     /product="nonfunctional neomycin phosphotransferase I"
                     /label=npt1
                     /note="truncated kanamycin-resistance determinant; derived
                     from bacterial transposon Tn903; expressed in Escherichia 
                     coli does not confer resistance to the aminoglycoside 
                     antibiotics neomycin and kanamycin"
                     /protein_id="ADV36645.2"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARKLLPFSPDSVVTHGD
                     FSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPD
                     MNKLQFHLMLDEFF"
     repeat_region   3080..3170
                     /note="telomeric repeat; derived from Cryptococcus
                     neoformans var. neoformans JEC21"
     regulatory      complement(3194..4129)
                     /note="Cryptococcal terminator; derived from region
                     downstream from the NMT1 (N-myristoyltransferase I) coding 
                     region in Cryptococcus neoformans var. grubii H99"
                     /regulatory_class="terminator"
     CDS             complement(4139..4930)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(4937..5769)
                     /note="ACT1 promoter; derived from a region upstream of the
                     ACT1 (actin) coding sequence in Cryptococcus neoformans 
                     var. grubii H99"
                     /regulatory_class="promoter"
     misc_feature    5810..7852
                     /note="RNA interference cassette; uses convergent promoters
                     to drive the transcription of an interfering construct in 
                     Cryptococcus neoformans"
     regulatory      complement(5810..6116)
                     /note="Gal7 (FNU16994) terminator; derived from a region 
                     downstream of the GAL7 (UDP glucose-D-galactose-1-phosphate
                     uridylyltransferase) coding sequence in C. neoformans var. 
                     neoformans JEC21"
                     /regulatory_class="terminator"
     regulatory      6140..6693
                     /note="Gal7 (FNU16994) promoter; derived from a region
                     upstream of the GAL7 (UDP glucose-D-galactose-1-phosphate 
                     uridylyltransferase) coding sequence in Cryptococcus 
                     neoformans var. neoformans JEC21."
                     /regulatory_class="promoter"
     misc_feature    6706..6957
                     /note="ura5 gene fragment; derived from the unspliced URA5 
                     (orotidine monophosphate pyrophosphorylase) coding sequence
                     in Cryptococcus neoformans var. neoformans JEC21; this 
                     fragment is not translated and functions only as a sentinel
                     for RNA interference; CPCURA5A"
     regulatory      complement(6964..7518)
                     /note="Gal7 (FNU16994) promoter; derived from a region
                     upstream of the GAL7 (UDP glucose-D-galactose-1-phosphate 
                     uridylyltransferase) coding sequence in Cryptococcus 
                     neoformans var. neoformans JEC21"
                     /regulatory_class="promoter"
     regulatory      7541..7847
                     /note="Gal7 (FNU16994) terminator; derived from a region 
                     downstream of the GAL7 (UDP glucose-D-galactose-1-phosphate
                     uridylyltransferase) coding sequence in Cryptococcus 
                     neoformans var. neoformans JEC21"
                     /regulatory_class="terminator"
     promoter        complement(7862..7880)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7887..7903)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      8045..8500
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"