LOCUS       Exported               13829 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pSCUDT2MFE6H, complete sequence.
ACCESSION   LT727633
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13829)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13829)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 13829)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13829
                     /organism="Yeast expression vector pSCUDT2MFE6H"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 2232."
                     /db_xref="taxon:1945503"
     misc_feature    1..6318
                     /label=2MU
                     /note="2MU"
     rep_origin      341..1683
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    673..720
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      703..1683
                     /label=2mu ori
                     /note="2mu ori"
     protein_bind    complement(3927..3974)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      6345..7326
                     /regulatory_class="promoter"
                     /note="CYC1"
     CDS             7350..7913
                     /codon_start=1
                     /note="unnamed protein product; mDHFR"
                     /protein_id="SJL89039.1"
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     regulatory      8002..8272
                     /regulatory_class="terminator"
                     /note="CYC1T"
     terminator      8022..8269
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        8281..8496
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             8497..9300
                     /codon_start=1
                     /note="unnamed protein product; URA3"
                     /protein_id="SJL89040.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(9439..9680)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(9462..10050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10221..11081)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL89041.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(11082..11186)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    complement(11320..11410)
                     /label=3UTR
                     /note="3UTR"
     CDS             complement(11411..12781)
                     /codon_start=1
                     /note="unnamed protein product; mIghg2b; fragment of heavy 
                     chain of mouse anti-hPLAP monoclonal antibody (E6H)"
                     /protein_id="SJL89042.1"
                     /translation="QVQLQQSGAELARPGASVKLSCKASGYTLTSYGISWVKQRTGQGL
                     EWIGEIYPGSGNSYFNEKFKGKATLTVDKSSSTAYLHLSSLTSEDSAVYFCAGPRQVGL
                     LPFGYWGQGTLVTASAAKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNS
                     GSLSSSVHTFPALLQSGLYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGP
                     TSTINPCPPCKECHKCPAPNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPD
                     VQISWFVNNVEVLTAQTQTHREDYNSTIRVVSALPIQHQDWMSGKEFKCKVNNKDLPAP
                     IERTISKIKGIVRAPQVYILSPPPEQLSRKDVSLTCLAVGFSPEDISVEWTSNGHTEEN
                     YKDTAPVLDSDGSYFIYSKLNMKTSKWEKTDSFSCNVRHEGLKNYYLKKTISRSPGK"
     CDS             complement(12785..13036)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=-alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDK"
     misc_feature    complement(12794..13036)
                     /label=ppMF
                     /note="ppMF"
     promoter        complement(13098..13539)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     regulatory      complement(13148..13531)
                     /regulatory_class="promoter"
                     /note="GAL1"
     protein_bind    13422..13539
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     regulatory      13552..13822
                     /regulatory_class="terminator"
                     /note="CYC1T"
     terminator      13572..13819
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"