pGREG505-T vector (V005886)

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LOCUS       40924_22661        9376 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PGREG expression vector pGREG505-T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9376)
  AUTHORS   Karreman RJ, Morrissey JP.
  TITLE     Controlling expression of genes in Saccharomyces cerevisiae using 
            variants of the pGREG recombinational vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9376)
  AUTHORS   Karreman RJ, Morrissey JP.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUN-2010) Microbiology Department, University College 
            Cork, College Rd, Cork City, Cork 0000, Ireland
REFERENCE   3  (bases 1 to 9376)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9376)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JUN-2010) Microbiology Department, University College Cork, 
            College Rd, Cork City, Cork 0000, Ireland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        762..1166
                     /label=LEU2 promoter
     CDS             1179..2270
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     misc_feature    complement(3005..3508)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        3545..3649
                     /label=AmpR promoter
     CDS             3650..4507
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4681..5269
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5557..5578
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5593..5623
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5631..5647
                     /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     5655..5671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5692..5710
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5729..6117
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     misc_feature    6155..6188
                     /note="Rec1 site; pGREG recombinational cloning site"
     misc_feature    6182..6187
                     /label=SalI restriction enzyme cut site
                     /note="SalI restriction enzyme cut site"
     promoter        6477..6664
                     /label=HIS3 promoter
     CDS             6665..7321
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     misc_feature    7350..7355
                     /label=SalI restriction enzyme cut site
                     /note="SalI restriction enzyme cut site"
     terminator      7362..7609
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    7622..7655
                     /note="LoxP site (cre/lox KanMX marker rescue); Guldener,
                     et. al. 1996, Nucleic Acids Research 24, 2519-2524"
     protein_bind    7622..7655
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(7713..9069)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     protein_bind    complement(9124..9157)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(9194..9212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9222..9238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"