LOCUS       Exported               10116 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pAS2.1-MLT5, complete sequence.
ACCESSION   LT726923
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10116)
  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  JOURNAL   Unpublished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 10116)
  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 10116)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10116
                     /organism="Yeast expression vector pAS2.1-MLT5"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5539."
                     /db_xref="taxon:1945447"
     rep_origin      2..1344
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(2..982)
                     /direction=LEFT
                     /label=2mu ori
                     /note="2mu ori"
     protein_bind    complement(965..1012)
                     /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      complement(1060..1164)
                     /regulatory_class="terminator"
                     /note="FLP terminator"
     promoter        1603..1883
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1884..2558
                     /codon_start=1
                     /note="unnamed protein product; TRP1"
                     /protein_id="SJL86953.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(2661..3116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     3261..3277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3312..3586)
                     /regulatory_class="terminator"
                     /note="CYC1 terminator"
     terminator      3314..3561
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3855..4304)
                     /codon_start=1
                     /note="unnamed protein product; CYH2"
                     /protein_id="SJL86954.1"
                     /translation="MPSRFTKTRKHRGHVSAGKGRIGKHRKHPGGRGMAGGQHHHRINM
                     DKYHPGYFGKVGMRYFHKQQAHFWKPVLNLDKLWTLIPEDKRDQYLKSASKETAPVIDT
                     LAAGYGKILGKGRIPNVPVIVKARFVSKLAEEKIRAAGGVVELIA"
     promoter        5072..5473
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      5075..5478
                     /regulatory_class="promoter"
                     /note="ADH1 promoter"
     CDS             5501..5941
                     /codon_start=1
                     /note="unnamed protein product; GALbd"
                     /protein_id="SJL86955.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             5993..6388
                     /codon_start=1
                     /note="unnamed protein product; hMLT fragment"
                     /protein_id="SJL86956.1"
                     /translation="MALLGDPLQALPPSAAPTGPLLAPPAGATLNRLREPLLRRLSELL
                     DQAPEGRGWRRLAELAGSRGRLRLSCLDLEQCSLKVLEPEGSPSLCLLKLMGEKGCTVT
                     ELSDFLQAMEHTEVLQLLSPPGIKITVN"
     misc_feature    5996..5998
                     /label=Kozak
                     /note="Kozak"
     regulatory      6416..6517
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     CDS             complement(6584..7792)
                     /codon_start=1
                     /note="unnamed protein product; IS10R"
                     /protein_id="SJL86957.1"
                     /translation="MCELDILHDSLYQFCPELHLKRLNSLTLACHALLDCKTLTLTELG
                     RNLPTKARTKHNIKRIDRLLGNRHLHKERLAVYRWHASFICSGNTMPIVLVDWSDIREQ
                     KRLMVLRASVALHGRSVTLYEKAFPLSEQCSKKAHDQFLADLASILPSNTTPLIVSDAG
                     FKVPWYKSVEKLGWYWLSRVRGKVQYADLGAENWKPISNLHDMSSSHSKTLGYKRLTKS
                     NPISCQILLYKSRSKGRKNQRSTRTHCHHPSPKIYSASAKEPWVLATNLPVEIRTPKQL
                     VNIYSKRMQIEETFRDLKSPAYGLGLRHSRTSSSERFDIMLLIALMLQLTCWLAGVHAQ
                     KQGWDKHFQANTVRNRNVLSTVRLGMEVLRHSGYTITREDLLVAATLLAQNLFTHGYAL
                     GKL"
     primer_bind     complement(7965..7981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(7970..8007)
                     /label=5 UTR of lacZ, incl. RBS
                     /note="5 UTR of lacZ, incl. RBS"
     protein_bind    7989..8005
                     /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)."
     regulatory      complement(8008..8091)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(8013..8043)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(8183..8585)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(8367..8955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9126..9986)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL86958.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9987..10091)
                     /gene="bla"
                     /label=AmpR promoter