pCORE6 vector (V008287)

Basic Vector Information

Vector Name:
pCORE6
Antibiotic Resistance:
Ampicillin
Length:
8237 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Chandran S, Noskov VN, Segall-Shapiro TH, Ma L, Whiteis C, Lartigue C, Jores J, Vashee S, Chuang R.
Promoter:
GAL1

pCORE6 vector Map

pCORE68237 bp400800120016002000240028003200360040004400480052005600600064006800720076008000HIS3 promoterHIS3f1 oriM13 fwdT7 promoterSK primerI-SceI restriction siteGAL1 promoterIntron-encoded endonuclease I-SceIUra3TEF promoterKanRKS primerT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterCEN/ARS

pCORE6 vector Sequence

LOCUS       V008287                 8237 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008287
VERSION     V008287
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8237)
  AUTHORS   Chandran S, Noskov VN, Segall-Shapiro TH, Ma L, Whiteis C, Lartigue
            C, Jores J, Vashee S, Chuang R.
  TITLE     TREC-IN: gene knock-in genetic tool for genomes cloned in yeast
  JOURNAL   BMC Genomics 15 (1), 1180 (2014)
   PUBMED   25539750
REFERENCE   2  (bases 1 to 8237)
  AUTHORS   Chuang R-Y., Noskov V, Ma L.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2014) Synthetic Biology, J. Craig Venter
            Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA
REFERENCE   3  (bases 1 to 8237)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8237)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "BMC
            Genomics"; date: "2014"; volume: "15"; issue: "1"; pages: "1180"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (17-DEC-2014) Synthetic Biology, J. Craig Venter Institute, 9704
            Medical Center Drive, Rockville, MD 20850, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        315..502
                     /label="HIS3 promoter"
     CDS             503..1159
                     /label="HIS3"
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2023..2039
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2046..2064
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2097..2113
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2118..2131
                     /label="I-SceI restriction site"
                     /note="I-SceI restriction site"
     promoter        2133..2574
                     /label="GAL1 promoter"
                     /note="inducible promoter, regulated by Gal4"
     CDS             2597..3301
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     CDS             complement(3379..4182)
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label="ura3"
                     /protein_id="AJD09819.1"
                     /translation="MSTKSYTSRAETHASPVASKLLRLMDEKKTNLCASLDVRSTDELL
                     KLVETLGPYICLLKTHVDILDDFSYEGTVVPLKALAEKYKFLIFEDRKFADIGNTVKLQ
                     YTSGVYRIAEWSDITNAHGVTGAGIVAGLKQGAQEVTKEPRGLLMLAELSSKGSLAHGE
                     YTKGTVDIAKSDKDFVIGFIAQNDMGGREEGFDWLIMTPGVGLDDKGDALGQQYRTVDE
                     VVSGGSDIIIVGRGLFAKGRDPKVEGERYRNAGWEAYQKRISAPH"
     gene            complement(3379..4182)
                     /gene="ura3"
                     /label="ura3"
                     /note="KlUra3"
     promoter        4498..4841
                     /label="TEF promoter"
                     /note="Ashbya gossypii TEF promoter"
     CDS             4882..5396
                     /codon_start=1
                     /gene="kanMX4"
                     /product="kanamycin resistance protein"
                     /label="kanMX4"
                     /note="KanR"
                     /protein_id="AJD09820.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGW"
     gene            4882..5396
                     /gene="kanMX4"
                     /label="kanMX4"
     primer_bind     complement(5417..5433)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(5463..5481)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5502..5518)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5526..5542)
                     /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(5550..5580)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5595..5616)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5904..6492)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6666..7523)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7524..7628)
                     /label="AmpR promoter"
     misc_feature    7665..8168
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"

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