LOCUS       Exported                5457 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Yeast dual vector pCtB1, complete sequence.
ACCESSION   KJ922019
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  AUTHORS   Lee WY, Chung S-C., Song J-Y., Cho KM.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUN-2014) BioMaterial Lab, Samsung Advanced Institute 
            of Technology, 130 Samsung-ro, Suwon 443803, Republic of Korea
REFERENCE   2  (bases 1 to 5457)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5457
                     /organism="Yeast dual vector pCtB1"
                     /mol_type="other DNA"
                     /db_xref="taxon:2042189"
     rep_origin      134..1298
                     /label=yeast 2micron replication origin
                     /note="yeast 2micron replication origin"
     primer_bind     1356..1372
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1379..1397
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1405..1513
                     /note="multi-cloning site derived from pBluescriptII SK(+)"
     misc_feature    1406..1513
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1423..1439
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1473..1489)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1523..1549
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1553..1799
                     /regulatory_class="terminator"
                     /note="TPS1 terminator; interior"
     gene            1801..3088
                     /gene="URA3"
                     /label=URA3
     promoter        1821..2036
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2037..2840
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /label=URA3
                     /protein_id="ATG88231.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     regulatory      2841..3088
                     /regulatory_class="terminator"
                     /gene="URA3"
                     /note="CYC1 terminator"
     terminator      2841..3088
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     regulatory      3089..3301
                     /regulatory_class="terminator"
                     /note="TPS1 terminator; lateral"
     promoter        complement(3315..3333)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3354..3370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3378..3394
                     /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(3402..3432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3447..3468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3740..4359
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      complement(3756..4344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(4514..5398)
                     /gene="Neo"
                     /label=Neo
     CDS             complement(4514..5329)
                     /codon_start=1
                     /gene="Neo"
                     /product="neomycin phosphotransferase"
                     /label=Neo
                     /protein_id="ATG88232.1"
                     /translation="MADVECTRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFR
                     LSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEV
                     PGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQD
                     DLDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA
                     DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4514..5305)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5330..5434)
                     /gene="bla"
                     /label=AmpR promoter