LOCUS       Exported                7589 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pSCUDMFE6L2, complete sequence.
ACCESSION   LT727450
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  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  JOURNAL   Unpublished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 7589)
  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 7589)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7589
                     /organism="Yeast expression vector pSCUDMFE6L2"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3205."
                     /db_xref="taxon:1945502"
     regulatory      27..1016
                     /regulatory_class="promoter"
                     /note="CYC1 promoter"
     CDS             1040..1603
                     /codon_start=1
                     /note="unnamed protein product; mDHFR"
                     /protein_id="SJL88494.1"
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     regulatory      1692..1962
                     /regulatory_class="terminator"
                     /note="CYC1 terminator"
     terminator      1712..1959
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        1978..2389
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     regulatory      1983..2388
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     CDS             2409..2789
                     /codon_start=1
                     /note="unnamed protein product; bleomycin"
                     /protein_id="SJL88495.1"
                     /translation="MTDQATPNLPSRDFDSTAAFYERLGFGIVFRDAGWMILQRGDLKL
                     EFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGTMA
                     ALVDPDGTLLRLIQNELLAGIS"
     regulatory      2888..3158
                     /regulatory_class="terminator"
                     /note="CYC1 terminator"
     terminator      2908..3155
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3164..3400)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3182..3770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3941..4801)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL88496.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4802..4906)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5021..5291)
                     /regulatory_class="terminator"
                     /note="CYC1 terminator"
     terminator      5024..5271
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    complement(5330..5479)
                     /label=3 UTR of mIghg2b
                     /note="3 UTR of mIghg2b"
     CDS             complement(5480..6124)
                     /codon_start=1
                     /note="unnamed protein product; mIghg2b; fragment of light 
                     chain of mouse anti-hPLAP monoclonal antibody (E6L)"
                     /protein_id="SJL88497.1"
                     /translation="DIQMTQSPASLSVSVGESVTITCRASENIYSNLAWYQQKQGKSPQ
                     LLVYVATKLVDGVPSRFSGSGSGTQYSLKINSLQSEDFGSYYCQHFWDTPFTFGSGTKL
                     EMKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSW
                     TDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC"
     CDS             complement(6128..6379)
                     /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"
     CDS             complement(6137..6379)
                     /codon_start=1
                     /note="unnamed protein product; ppMF"
                     /protein_id="SJL88498.1"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVS"
     promoter        complement(6441..6882)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     regulatory      complement(6491..6882)
                     /regulatory_class="promoter"
                     /note="GAL1 promoter"
     protein_bind    6765..6882
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     misc_feature    complement(6829..6845)
                     /label=UAS
                     /note="UAS"
     misc_feature    complement(6847..6863)
                     /label=UAS
                     /note="UAS"
     misc_feature    complement(6866..6882)
                     /label=UAS
                     /note="UAS"
     gap             6892..7192
                     /estimated_length=301
     misc_feature    7223..7561
                     /label=Ty1
                     /note="Ty1"