pFA-URA3-Clox vector (V015758)

Basic Vector Information

Vector Name:
pFA-URA3-Clox
Antibiotic Resistance:
Ampicillin
Length:
6763 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F

pFA-URA3-Clox vector Map

pFA-URA3-Clox6763 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600SK primerloxPOrotidine 5'-phosphate decarboxylaseCaMET3 promoter; inducible upon growth without methionine repressible via addition of methionine and cysteine to the mediumCre3'UTRderived from Saccharomyces cerevisiae CYC1loxPT7 promoteroriAmpRAmpR promoterSP6 promoter

pFA-URA3-Clox vector Sequence

LOCUS       V015758                 6763 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V015758
VERSION     V015758
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6763)
  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 6763)
  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 6763)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene v. 13
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE
            (2019) In press"
            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"
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..69
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    83..116
                     /label="loxP"
                     /note="loxP"
     CDS             546..1355
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      1500..2831
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(2859..3262,3427..4054)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59752.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          3263..3426
                     /note="modified TUB2 intron sequence"
     3'UTR           4067..4197
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4198..4257
                     /label="derived from Saccharomyces cerevisiae CYC1"
                     /note="derived from Saccharomyces cerevisiae CYC1"
                     /regulatory_class="terminator"
     protein_bind    complement(4270..4303)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4401..4419)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(4677..5265)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5439..6296)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6297..6401)
                     /label="AmpR promoter"
     promoter        6747..6763
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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