LOCUS       Exported                8237 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCORE6, complete sequence.
ACCESSION   KP282615
VERSION     .
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8237
                     /organism="Cloning vector pCORE6"
                     /mol_type="other DNA"
                     /db_xref="taxon:1592726"
     promoter        315..502
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             503..1162
                     /codon_start=1
                     /gene="his3"
                     /product="His3"
                     /label=his3
                     /protein_id="AJD09818.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     gene            503..1162
                     /gene="his3"
                     /label=his3
     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..5396
                     /label=CORE6 cassette
                     /note="CORE6 cassette"
     misc_feature    2118..2131
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     regulatory      2132..2590
                     /regulatory_class="promoter"
                     /note="Gal1"
     promoter        2133..2574
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2133..2250
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             2591..3304
                     /codon_start=1
                     /gene="I-SceI"
                     /product="I-SceI"
                     /label=I-SceI
                     /note="restriction enzyme"
                     /protein_id="AJD09817.1"
                     /translation="MHMKNIKKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILG
                     DAYIRSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKERVNHLGNLVITWGAQ
                     TFKHQAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIV
                     LNTQSFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQM
                     MYKLPNTISSETFLK"
     gene            2591..3304
                     /gene="I-SceI"
                     /label=I-SceI
     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"
     5UTR           4498..4841
                     /label=translation elongation factor 1 alpha
                     /note="translation elongation factor 1 alpha"
     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    5526..5542
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
     rep_origin      complement(5904..6492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6663..7523)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7524..7628)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7665..8168
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"