LOCUS       Exported                6449 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJG4-5 (pB42AD), complete sequence.
ACCESSION   U89961
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  AUTHORS   Gyuris J, Golemis E, Chertkov H, Brent R.
  TITLE     Cdi1, a human G1 and S phase protein phosphatase that associates 
            with Cdk2
  JOURNAL   Cell 75 (4), 791-803 (1993)
  PUBMED    8242750
REFERENCE   2  (bases 1 to 6449)
  AUTHORS   Gyuris J, Golemis E, Chertkov H, Brent R.
  TITLE     pB42AD (pJG4-5) complete sequence
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6449)
  AUTHORS   Golemis E, Gyuris J, Chertkov H, Brent R.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-FEB-1997) CLONTECH Laboratories, Inc., 1020 East 
            Meadow Circle, Palo Alto, CA 94303-4230, USA
REFERENCE   4  (bases 1 to 6449)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     This vector can be obtained from CLONTECH Laboratories, Inc., 1020 
            East Meadow Circle, Palo Alto, CA 94303-4230, USA. To place an order
            call (415) 424-8222 or (800) 662-2566, extension 1. International 
            customers, please contact your local distributor. For technical 
            information, call (415) 424-8222 or (800) 662-2566, extension 3.
            This sequence has been compiled from information in the sequence 
            databases, published literature and other sources, together with 
            partial sequences obtained by CLONTECH. If you suspect there is an 
            error in this sequence, please contact CLONTECHs Technical Support 
            Department at (415) 424-8222 or (800) 662-2566, extension 3 or 
            E-mail TECH@CLONTECH.COM.
FEATURES             Location/Qualifiers
     source          1..6449
                     /organism="Cloning vector pJG4-5"
                     /mol_type="genomic DNA"
                     /label=commercially available as pB42AD
                     /note="commercially available as pB42AD"
                     /db_xref="taxon:57583"
     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"
     CDS             534..845
                     /codon_start=1
                     /product="B42 transcription activation domain-HA1 epitope 
                     tag fusion protein"
                     /label=B42 transcription activation domain-HA1 epitope
                     /protein_id="AAB68652.1"
                     /translation="MGAPPKKKRKVAGINKDIEECNAIIEQFIDYLRTGQEMPMEMADQ
                     AINVVPGMTPKTILHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGDASYPYDVPDYAS
                     "
     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_feature    817..845
                     /label=encodes HA1 epitope tag
                     /note="encodes HA1 epitope tag"
     terminator      1004..1191
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      1713..3055
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    2045..2092
                     /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)."
     mRNA            complement(3365..4190)
     CDS             complement(3416..4090)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(4091..4192)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        4295..4399
                     /gene="bla"
                     /label=AmpR promoter
     gene            4400..5261
                     /gene="ampR"
                     /label=ampR
     CDS             4400..5260
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=ampR
                     /protein_id="AAB68653.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6307..6328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6343..6373
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6381..6397
                     /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     6405..6421
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"