CIpNAT plasmid CIpNat vector (V010036)

Basic Vector Information

      • Vector Name:
      • CIpNAT plasmid CIpNat
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7822 bp
      • Type:
      • Integrating vector
      • Source/Author:
      • Shahana S, Childers DS, Ballou ER, Bohovych I, Odds FC, Gow NA, Brown AJ.

CIpNAT plasmid CIpNat vector Vector Map

CIpNAT plasmid CIpNat7822 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800f1 orim13fT7RPS1 targeting sequenceSK primerFLP recombination target sequence (FRT) (Staib et al., 1999)Ashbya gossypii TEF1 promoter (Shen et al., 2005)CaNatAshbya gossypii TEF1 terminator (Shen et al., 2005)FLP1Candida albicans SAP2 promoter (Staib et al., 1999)SK primerFLP recombination target sequence (FRT) (Staib et al., 1999)T3m13rlac operatorlac promoterCAP binding siteColE1 oriAmpRAmpR 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 ℃.

CIpNAT plasmid CIpNat vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7822 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integrating vector CIpNAT plasmid CIpNat, complete sequence.
ACCESSION   KJ174065
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7822)
  AUTHORS   Shahana S, Childers DS, Ballou ER, Bohovych I, Odds FC, Gow NA, 
            Brown AJ.
  TITLE     New Clox Systems for Rapid and Efficient Gene Disruption in Candida 
            albicans
  JOURNAL   PLoS ONE 9 (6), E100390 (2014)
  PUBMED    24940603
REFERENCE   2  (bases 1 to 7822)
  AUTHORS   Shahana S, Childers DS, Ballou ER, Bohovych I, Odds FC, Gow NA, 
            Brown AJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2014) School of Medical Sciences, University of 
            Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25
            2ZD, UK
REFERENCE   3  (bases 1 to 7822)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7822
                     /organism="Integrating vector CIpNAT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1516111"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(12..452)
                     /direction=LEFT
                     /label=F1 ori
                     /note="F1 ori"
     misc_feature    602..619
                     /label=m13f
                     /note="m13f"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    629..648
                     /label=T7
                     /note="T7"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_recomb     1040..1513
                     /label=RPS1 targeting sequence
                     /note="RPS1 targeting sequence"
     primer_bind     complement(1511..1527)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1544..1560
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1562..1595)
                     /note="FLP recombination target sequence (FRT) (Staib et 
                     al., 1999)"
     protein_bind    1562..1595
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      1597..1979
                     /regulatory_class="promoter"
                     /gene="TEF1"
                     /note="Ashbya gossypii TEF1 promoter (Shen et al., 2005)"
     gene            1597..1979
                     /gene="TEF1"
                     /label=TEF1
     promoter        1636..1979
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1993..2565
                     /codon_start=1
                     /gene="CaNat"
                     /product="CaNat"
                     /label=CaNat
                     /note="nat1 from Streptomyces noursei; codon optimized for 
                     Candida albicans by Shen et al., 2005"
                     /protein_id="AIE16102.1"
                     /translation="MSTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     gene            1993..2565
                     /gene="CaNat"
                     /label=CaNat
     regulatory      2575..2815
                     /regulatory_class="terminator"
                     /note="Ashbya gossypii TEF1 terminator (Shen et al., 2005)"
     terminator      2580..2777
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     regulatory      complement(2810..3206)
                     /regulatory_class="terminator"
                     /note="Candida albicans ACT1 terminator (Staib et al., 
                     1999)"
     CDS             complement(3204..4475)
                     /codon_start=1
                     /gene="FLP1"
                     /product="FLP1"
                     /label=FLP1
                     /note="Saccharomyces cerevisiae FLP1 recombinase codon 
                     optimized for Candida albicans (Staib et al., 1999)"
                     /protein_id="AIE16103.1"
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     gene            complement(3204..4475)
                     /gene="FLP1"
                     /label=FLP1
     regulatory      complement(4500..5542)
                     /regulatory_class="promoter"
                     /gene="SAP2"
                     /note="Candida albicans SAP2 promoter (Staib et al., 1999)"
     gene            complement(4500..5542)
                     /gene="SAP2"
                     /label=SAP2
     primer_bind     5554..5570
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(5572..5605)
                     /note="FLP recombination target sequence (FRT) (Staib et 
                     al., 1999)"
     protein_bind    5572..5605
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    complement(5636..5655)
                     /label=T3
                     /note="T3"
     promoter        complement(5638..5656)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    5673..5693
                     /label=m13r
                     /note="m13r"
     primer_bind     complement(5677..5693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5701..5717
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5724..5754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5769..5790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6060..6688)
                     /direction=LEFT
                     /label=ColE1 ori
                     /note="ColE1 ori"
     rep_origin      complement(6078..6666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6837..7697)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="ampicillin resistance; pBluescript KS+ backbone 
                     (Staib et al., 1999)"
                     /protein_id="AIE16104.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6837..7697)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(7698..7802)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(7739..7767)
                     /regulatory_class="promoter"
                     /gene="amp"
                     /note="pBluescript KS+ backbone (Staib et al., 1999)"
     gene            complement(7739..7767)
                     /gene="amp"
                     /label=amp

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