pGEM-T_blanc_GFPco8g_tGAL_URA3locus_tTK vector (V016063)

LOCUS       62056_11390        7148 bp DNA     circular SYN 23-MAR-2020
DEFINITION  Cloning vector pGEM-T_blanc_GFPco8g_tGAL_URA3locus_tTK, complete 
            sequence.
ACCESSION   MN271957
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  AUTHORS   Lodens S, Roelants SLK.W., Geys R, Coussement P, De Maeseneire SL, 
            Soetaert W.
  TITLE     The effect of its gene cluster localization on sophorolipid 
            production by Starmerella bombicola
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7148)
  AUTHORS   Lodens S, Roelants SLK.W.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-AUG-2019) Inbio.be; Department of Biotechnology, Ghent
            University, Coupure links 653, Ghent 9000, Belgium
REFERENCE   3  (bases 1 to 7148)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-AUG-2019) Inbio.be; Department of Biotechnology, Ghent 
            University, Coupure links 653, Ghent 9000, Belgium"
FEATURES             Location/Qualifiers
     source          1..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    53..1843
                     /label=5' homologous region ura3 locus
                     /note="5' homologous region ura3 locus"
     regulatory      427..1054
                     /regulatory_class="promoter"
     gene            1055..1843
                     /gene="ura3"
                     /label=ura3
     CDS             1055..1843
                     /codon_start=1
                     /transl_table=11
                     /gene="ura3"
                     /product="URA3"
                     /label=ura3
                     /protein_id="QIM58711.1"
                     /translation="MKLSYEERRERLPAVAARLFDIIVSKQTNLCASLDVRTTSELLSI
                     LDRIGPYICMVKTHIDIIDDFEYDTTVSGLKQLSTKHNFLIFEDRKFADIGSTVKAQYA
                     GGVFKIAQWADITNAHGVPGPGIVSGLEEAAKETTDEPRGLVMLAELSSKGTLAHGEYS
                     QATVDIARSNRAFVFGFIAQQKVGKPEEDWVIMTPGVGLDDKGDGLGQQYRTVDDVIET
                     GTDVIIVGRGLYSKGRDPVHEAQRYQKAGWNAYLRKVQSR"
     polyA_signal    1908..1956
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             1976..2689
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      2693..3161
                     /regulatory_class="terminator"
     misc_feature    3162..4210
                     /label=3' homologous region ura3 locus
                     /note="3' homologous region ura3 locus"
     promoter        complement(4273..4291)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4309..4325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4333..4349)
                     /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)."
     promoter        complement(4357..4387)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4402..4423)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4711..5299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5473..6330)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6331..6435)
                     /label=AmpR promoter
     rep_origin      complement(6513..6968)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7109..7125
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7132..7148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"