pDIS-NAT1-Clox vector (V015349)

Basic Vector Information

Vector Name:
pDIS-NAT1-Clox
Antibiotic Resistance:
Ampicillin
Length:
9771 bp
Type:
Cloning vector
Replication origin:
ori
Host:
Yeast
Source/Author:
Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F
Promoter:
TEF

pDIS-NAT1-Clox vector Map

pDIS-NAT1-Clox9771 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600NEUT5L 3'M13 fwdT7 promoterSK primerloxPTEF terminatorNrsRTEF promoterCaMET3 promoter; inducible upon growth without methionine repressible via addition of methionine and cysteine to the mediumCre3'UTRTranscriptional termination from Saccharomyces cerevisiae CYC1 geneloxPKS primerNEUT5L 5'lac promoterCAP binding siteoriAmpRAmpR promoterCEN/ARSTRP1 promoterTRP1

pDIS-NAT1-Clox vector Sequence

LOCUS       62056_8465        9771 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pDIS-NAT1-Clox, complete sequence.
ACCESSION   MK652134
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9771)
  AUTHORS   Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F, 
            Correa-Bordes J, Vazquez de Aldana CR.
  TITLE     A new toolkit for gene tagging in Candida albicans containing 
            recyclable markers
  JOURNAL   PLoS ONE (2019) In press
REFERENCE   2  (bases 1 to 9771)
  AUTHORS   Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F, 
            Correa-Bordes J, Vazquez de Aldana CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2019) Instituto de Biologia Funcional y Genomica 
            (IBFG), Consejo Superior de Investigaciones Cientificas, Zacarias 
            Gonzalez 2, Salamanca, Salamanca 37007, Spain
REFERENCE   3  (bases 1 to 9771)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE 
            (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-MAR-2019) Instituto de Biologia Funcional y Genomica (IBFG), 
            Consejo Superior de Investigaciones Cientificas, Zacarias Gonzalez 
            2, Salamanca, Salamanca 37007, Spain"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene v. 13
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(166..465)
                     /label=NEUT5L 3'
                     /note="NEUT5L 3'"
     primer_bind     736..752
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        759..777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     817..833
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    847..880
                     /label=loxP
                     /note="loxP"
     terminator      complement(1291..1488)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1506..2069)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(2089..2432)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2496..3827
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3855..4258,4423..5050)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59800.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4259..4422
                     /note="modified TUB2 intron sequence"
     3'UTR           5063..5139
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5194..5253
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5266..5299)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5379..5395)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(5406..5655)
                     /label=NEUT5L 5'
                     /note="NEUT5L 5'"
     promoter        complement(5670..5700)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5715..5736)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6024..6612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6786..7643)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7644..7748)
                     /label=AmpR promoter
     misc_feature    7785..8288
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        8544..8824
                     /label=TRP1 promoter
     CDS             8825..9496
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"

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