LOCUS       Exported                8718 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEZY45, complete sequence.
ACCESSION   EU181460
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8718)
  AUTHORS   Guo F, Wang Y, Zhang YZ.
  TITLE     Construction of two recombination yeast two-hybrid vectors by in 
            vitro recombination
  JOURNAL   Mol. Biotechnol. 36 (1), 38-43 (2007)
  PUBMED    17827536
REFERENCE   2  (bases 1 to 8718)
  AUTHORS   Guo F, Wang Y, Zhang Y-Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2007) Biological, Chemical, and Physical Sciences,
            Illinois Institute of Technology, 3101 South Dearborn St., Chicago, 
            IL 60616, USA
REFERENCE   3  (bases 1 to 8718)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8718
                     /organism="Cloning vector pEZY45"
                     /mol_type="other DNA"
                     /label=yeast two-hybrid vector
                     /note="yeast two-hybrid vector"
                     /db_xref="taxon:474434"
     regulatory      1..523
                     /regulatory_class="promoter"
                     /note="GAL1 promoter"
     promoter        81..522
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    81..198
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     misc_feature    534..848
                     /note="encodes partial B42 transcription activation 
                     domain/HA1 epitope tag fusion protein"
     CDS             546..566
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             573..809
                     /codon_start=1
                     /product="yeast transcriptional activator cretaed from E. 
                     coli genomic DNA fragments (Ma and Ptashne, 1987)"
                     /label=B42 transcriptional activator
                     /translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI
                     LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD"
     CDS             816..842
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_recomb     861..986
                     /label=attR1 site
                     /note="attR1 site"
     protein_bind    861..985
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1414..1719)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB lethal protein"
                     /function="gyrase inhibitor"
                     /label=ccdB
                     /protein_id="ABW37353.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(1414..1719)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(2064..2723)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /protein_id="ABW37354.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(2064..2723)
                     /gene="CmR"
                     /label=CmR
     promoter        complement(2724..2826)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_recomb     3004..3129
                     /label=attR2 site
                     /note="attR2 site"
     protein_bind    complement(3004..3128)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3273..3460
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      3305..3599
                     /regulatory_class="terminator"
                     /note="ADH terminator"
     rep_origin      3744..5639
                     /label=2 mu ori
                     /note="2 mu ori"
     rep_origin      3982..5324
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    4314..4361
                     /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)."
     CDS             complement(5685..6359)
                     /codon_start=1
                     /gene="Trp1"
                     /product="Trp1"
                     /label=Trp1
                     /protein_id="ABW37355.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     gene            complement(5685..6359)
                     /gene="Trp1"
                     /label=Trp1
     promoter        complement(6360..6461)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        6564..6668
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6669..7529
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicillin resistance protein; AmpR"
                     /protein_id="ABW37356.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            6669..7529
                     /gene="ampR"
                     /label=ampR
     rep_origin      7674..8347
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      7700..8288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8576..8597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8612..8642
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8650..8666
                     /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)."
     primer_bind     8674..8690
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"