pLNMCL vector (V004964)

LOCUS       40924_28556       10495 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLNMCL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10495)
  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 10495)
  AUTHORS   Shahana S, Childers DS, Brown AJP.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-MAY-2013) Microbiology, School of Medical Sciences, 
            Institute of Medical Sciences - Foresterhill, Aberdeen, 
            Aberdeenshire AB25 2ZD, United Kingdom
REFERENCE   3  (bases 1 to 10495)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10495)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2014"; volume: "9"; issue: "6"; pages: "E100390"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-MAY-2013) Microbiology, School of Medical Sciences, Institute of
            Medical Sciences - Foresterhill, Aberdeen, Aberdeenshire AB25 2ZD, 
            United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(666..1757)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(1770..2174)
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3097..3115
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3148..3164
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3178..3211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      complement(3622..3819)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(3837..4400)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(4420..4763)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      4827..6158
                     /gene="pMET3"
                     /label=MET3 promoter
                     /note="MET3 promoter"
                     /regulatory_class="promoter"
     gene            4827..6158
                     /gene="pMET3"
                     /label=pMET3
     CDS             6186..6593
                     /codon_start=1
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="AGZ63435.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQAL"
     protein_bind    complement(7597..7630)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7675..7691)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7721..7739)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7760..7776)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7784..7800
                     /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(7808..7838)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7853..7874)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8162..8750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8924..9781)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9782..9886)
                     /label=AmpR promoter
     misc_feature    9923..10426
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"