pPL5617_pUG_PuroR vector (V004045)

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LOCUS       40924_34889        3780 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPL5617_pUG_PuroR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3780)
  AUTHORS   MacDonald C, Piper RC.
  TITLE     Puromycin- and methotrexate-resistance cassettes and optimized 
            Cre-recombinase expression plasmids for use in yeast
  JOURNAL   Yeast 32 (5), 423-438 (2015)
  PUBMED    25688547
REFERENCE   2  (bases 1 to 3780)
  AUTHORS   MacDonald C, Piper RC.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 3780)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3780)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2015"; volume: "32"; issue: "5"; pages: "423-438"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-NOV-2014) Molecular Physiology "
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Sequence Builder (Lasergene) v. 8.0.3 
            Sequencing Technology :: Assembled
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             485..1081
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     terminator      1090..1287
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1331..1364
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1331..1364)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1418..1436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1694..2282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2456..3313)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3314..3418)
                     /label=AmpR promoter
     promoter        3764..3780
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"