pFA-HAH2 vector (V006383)

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LOCUS       40924_19111        8618 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Functional analysis vector pFA-HAH2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8618)
  AUTHORS   Sharma S, Alfatah M, Bari VK, Rawal Y, Paul S, Ganesan K.
  TITLE     Sphingolipid Biosynthetic Pathway Genes FEN1 and SUR4 Modulate 
            Amphotericin B Resistance
  JOURNAL   Antimicrob. Agents Chemother. 58 (4), 2409-2414 (2014)
  PUBMED    24395234
REFERENCE   2  (bases 1 to 8618)
  AUTHORS   Paul S, Rawal Y, Alfatah M, Bari VK, Sharma S, Choudhury S, Malik S,
            Ganesan K.
  TITLE     Single-Transformation Gene Function Test by Conditional Expression 
            of Both Alleles of Candida albicans Genes
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 8618)
  AUTHORS   Paul S, Rawal Y, Alfatah M, Bari VK, Sharma S, Choudhury S, Malik S,
            Ganesan K.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JUL-2013) Institute of Microbial Technology, Sector 
            39-A, Chandigarh, UT 160036, India
REFERENCE   4  (bases 1 to 8618)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8618)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob.
            Agents Chemother."; date: "2014"; volume: "58"; issue: "4"; pages: 
            "2409-2414"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (03-JUL-2013) Institute of Microbial Technology, Sector 39-A, 
            Chandigarh, UT 160036, India"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..55
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            60..928
                     /gene="HIS1"
                     /label=HIS1
                     /note="5' region; derived from Candida albicans HIS1"
     repeat_region   575..928
                     /gene="HIS1"
     gene            934..2884
                     /gene="ARG4"
                     /label=ARG4
                     /note="derived from Candida albicans ARG4"
     CDS             1391..2797
                     /codon_start=1
                     /gene="ARG4"
                     /product="arg4"
                     /label=ARG4
                     /note="argininosuccinate lyase"
                     /protein_id="AGY46360.1"
                     /translation="MSQQQDKQPSENKLWGGRFTGATDPLMDLYNASLPYDKVMYDADL
                     TGTKVYTQGLNKLGLITTEELNLIHQGLEQIRQEWHDNKFIIKAGDEDIHTANERRLGE
                     IIGKNISGKVHTGRSRNDQVATDMRIFVRESLLNLSKILHQFITAILERAHKEIDVLMP
                     GYTHLQKAQPIRWAHWLSSYATYFTEDYKRLQEIITRVNQSPLGSGALAGHPYGIDREF
                     LAKGLGFDGVIGNSLTAVSDRDFVVESLFWSTLFMNHISRFSEDLIIYSSGEFGFIKLA
                     DAYSTGSSLMPQKKNPDSLELLRGKSGRVFGQLSGFLMSIKSIPSTYNKDMQEDKEPLF
                     DALTTVEHSILIATGVISTLLIDKQNMEKALTMDMLATDLADYLVRKGVPFRETHHISG
                     ECVRKAEEEKLSGIDQLSFEQFQQIDSRFEKDVMETFDFEASVERRDALGGTAKSAVLK
                     QLENLKSILS"
     gene            2889..3738
                     /gene="HIS1"
                     /label=HIS1
                     /note="3' region; derived from Candida albicans HIS1"
     repeat_region   2889..3242
                     /gene="HIS1"
     protein_bind    3739..3772
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(3831..3849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3859..3875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4020..4475
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4609..5409)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5410..5630)
                     /label=URA3 promoter
     misc_feature    complement(5895..6398)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        6435..6539
                     /label=AmpR promoter
     CDS             6540..7397
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7571..8159
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8447..8468
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8483..8513
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8521..8537
                     /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     8545..8561
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8582..8600
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"