pBM5155 vector (V009142)

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LOCUS       40924_6831        9405 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBM5155, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9405)
  AUTHORS   Alexander WG, Peris D, Pfannenstiel BT, Opulente DA, Kuang M, 
            Hittinger CT.
  TITLE     Efficient engineering of marker-free synthetic allotetraploids of 
            Saccharomyces
  JOURNAL   Fungal Genet. Biol. (2015) In press
  PUBMED    26555931
REFERENCE   2  (bases 1 to 9405)
  AUTHORS   Alexander WG, Peris D, Pfannenstiel B, Opulente DA, Kuang M, 
            Hittinger CT.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-SEP-2015) Laboratory of Genetics, University of 
            Wisconsin-Madison, 425-G Henry Mall, 4350 Genetics Biotechnology 
            Center, Madison, WI 53706, USA
REFERENCE   3  (bases 1 to 9405)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9405)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal 
            Genet. Biol. (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-SEP-2015) Laboratory of Genetics, University of 
            Wisconsin-Madison, 425-G Henry Mall, 4350 Genetics Biotechnology 
            Center, Madison, WI 53706, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(31..1035)
                     /codon_start=1
                     /label=tTA
                     /note="tetracycline-controlled transactivator, comprising a
                     fusion of the tetracycline repressor TetR with the 
                     C-terminal activation domain of herpes simplex virus VP16"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     promoter        complement(1140..1343)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(1344..1723)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     terminator      1835..2022
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     protein_bind    2052..2322
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     misc_feature    2540..2547
                     /label=NotI site for gap-repair cloning
                     /note="NotI site for gap-repair cloning"
     misc_feature    2585..2608
                     /label=6x glycine linker
                     /note="6x glycine linker"
     CDS             2609..3106
                     /codon_start=1
                     /label=13xMyc
                     /note="13 tandem Myc epitope tags"
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     misc_feature    3251..3253
                     /label=in-frame amber stop codon
                     /note="in-frame amber stop codon"
     terminator      3299..3546
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3569..3585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3593..3609)
                     /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(3617..3647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3662..3683)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3971..4559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4733..5590)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5591..5695)
                     /label=AmpR promoter
     gene            5903..7022
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     rep_origin      7365..8202
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     misc_feature    8338..8630
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     primer_bind     9389..9405
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"