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ID PGAD424 preliminary; circular DNA; SYN; 6659 BP.
XX
AC U07647;
XX
DT 01-JUL-1994 (Rel. 9, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE Yeast/E.coli plasmid vector pGAD424 - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RP 1-6659
RC pGAD424 from ADH1 promoter & GAL4 & ADH1 terminator & LEU2 gene
RA Kitts P.A., Ledesma L.J.;
RT ;
RL Unpublished (1994).
XX
RN [2]
RP 1-6659
RC pBTM116 from ADH1 promoter/terminator & TRP1 gene & lexA gene
RC pGBT9 from ADH1 promoter/terminator & GAL4 & TRP1 gene
RC pGAD10 from ColE1 & GAL4 & LEU2 gene
RC pGAD424 from ADH1 promoter/terminator & GAL4 & LEU2 gene
RC [lambda ACT from lambda & LEU2 gene]
RA Bartel P.L., Chien C., Sternglanz R., Fields S.;
RT "Using the two-hybrid system to detect protein-protein interactions";
RL Cellular Interactions In Development 0:153-179(1993).
RL Hartley D., ed., Oxford University Press, Oxford.
XX
RN [3]
RP 1-6659
RC pGAD424
RA Kitts P.A.;
RT ;
RL Submitted (11-MAR-1994) by:
RL Kitts P.A., CLONTECH Laboratories, Inc.,
RL 4030 Fabian Way, Palo Alto, CA 94303, USA.
XX
RN [4]
RC from pGAD424
RC from pGBT9
RA Bartel P.L., Zhu L.;
RT "Rapid Identification of genes encoding interacting proteins with
RT the MATCHMAKER[tm] two-hybrid system";
RL CLONTECHniques VIII4:1-5(1993).
XX
RN [5]
RC from pGAD424
RC from pGBT9
RA Hannon G.J., Zhu L., Holtz A.;
RT "Screening for interacting proteins using the MATCHMAKER[tm]
RT two-hybrid system";
RL CLONTECHniques IX1:1-4(1994).
XX
RN [6]
RC [from none]
RA Fields S., Sternglanz R.;
RT "The two-hybrid system: an assay for protein-protein interactions";
RL Trends Genet. 10:286-292(1994).
XX
RN [7]
RC pGX13 from pKLC15 & 401-N
RC pAE7 from pAAH5
RC pGAD1, pGAD2, pGAD3 from pGX13 & pAE7
RC pGAD1F, pGAD2F, pGAD3F from pGX13 & pAE7
RC plasmid from oligo & pES15, SIR1 gene
RC pKL5 from pMA424 & plasmid
RC pCTC7 from pMA424 & pJM100, SIR2 gene
RC pKL3 from pUC18 & pJR104, SIR3 gene
RC pKL13 from pKL3 & pGAD3
RC pCTC15 from pUC18 & pJR643, SIR4 gene
RC pCTC17, pCTC18 from pCTC15 & pMA424
RC pCTC23 from pMA424 & pJR643
RC pCTC24 from pGAD2 & pJR643
RC pCTC14 from pGAD3 & pSL321, SNF4 gene
RC pTD1 from pGAD3F & SV40 large T antigen
RA Chien C.T., Bartel P.L., Sternglanz R., Fields S.;
RT "The two-hybrid system: a method to identify and clone genes for
RT proteins that interact with a protein of interest";
RL Proc. Natl. Acad. Sci. U.S.A. 88:9578-9582(1991).
XX
RN [8]
RC GAL4(1-881), GAL4(1-147) from GAL4 gene
RC GAL4(1-147)-SNF1 from GAL4 gene & SNF1 gene
RC SNF4-GAL4(768-881) from GAL4 gene & SNF4 gene
RC pCL1 from YCp50 & GAL4 gene
RA Fields S., Song O.;
RT "A novel genetic system to detect protein-protein interactions";
RL Nature 340:245-247(1989).
XX
RN [9]
RC from pGAD424
RC from pGAD1F
RC from pGAD-GH
RC from pACT
RC from pPC86
RA Fields S.;
RT ;
RL METHODS A Companion to Methods in Enzymology 5:116-124(1993).
XX
RN [10]
RC [from none]
RA Fritz C.C., Green M.R.;
RT ;
RL Curr. Biol. 2:403-405(1992).
XX
RN [11]
RC [from none]
RA Guarente L.;
RT "Strategies for the identification of interacting proteins";
RL Proc. Natl. Acad. Sci. U.S.A. 90:1639-1641(1993).
XX
RN [12]
RC pRY121-2micron- from pRY121
RC pCZIGAL from pCZ3GAL
RC pVA1, pVA3 from pMA424 & mouse p53 gene
RC pLAM5 from pMA424 & human lamin C protein gene
RC pTD1 from pGAD3F & SV40 large T antigen
RA Bartel P.L., Chien C.T., Sternglanz R., Fields S.;
RT "Elimination of false positives that arise in using the two-hybrid
RT system";
RL Biotechniques 14:920-924(1993).
XX
RN [13]
RC from YEplac195
RC from YCplac33
RA Gietz D., St Jean A., Woods R.A., Schiestl R.H.;
RT "Improved method for high efficiency transformation of intact
RT yeast cells";
RL Nucleic Acids Res. 20:1425-1425(1992).
XX
RN [14]
RC from YEp351
RC from YCp50
RC NOTE: see Nucleic Acids Res. 19:6688-6688(1991)
RA Hill J., Donald K.A., Griffiths D.E.;
RT "DMSO-enhanced whole cell yeast transformation";
RL Nucleic Acids Res. 19:5791-5791(1991).
XX
RN [15]
RC from YRp7
RC from YEp6
RC from pDB248
RA Ito H., Fukuda Y., Murata K., Kimura A.;
RT "Transformation of intact yeast cells treated with alkali cations";
RL J. Bacteriol. 153:163-168(1983).
XX
RN [16]
RC from YEplac195
RC from YEplac181
RC from pYAC3
RC from pDG95
RA Schiestl R.H., Gietz R.D.;
RT "High efficiency transformation of intact yeast cells using single
RT stranded nucleic acids as a carrier";
RL Curr. Genet. 16:339-346(1989).
XX
RN [17]
RC from pGBT9, GAL4 gene
RC from pCL1
RC from pGAD-GH
RC pGBT9-SNF1 from pGBT9 & SNF1 gene
RC pGADGH-SNF4 from pGAD-GH & SNF4 gene
RA Feilotter H.E., Hannon G.J., Ruddell C.J., Beach D.;
RT "Construction of an improved host strain for two hybrid screening";
RL Nucleic Acids Res. 22:1502-1503(1994).
XX
RN [18]
RC pGAD-GH from pGAD series
RA Hannon G.J., Demetrick D., Beach D.;
RT "Isolation of the Rb-related p130 through its interaction with
RT CDK2 and cyclins";
RL Genes Dev. 7:2378-2391(1993).
XX
RN [19]
RC pES15 from pUC18 & SIR1 gene
RA Stone E.;
RT ;
RL Unpublished (1991).
XX
RN [20]
RC 401-N from SV40 & T7
RA Benton B.M., Eng W.K., Dunn J.J., Studier F.W., Sternglanz R.,
RA Fisher P.A.;
RT "Signal-mediated import of bacteriophage T7 RNA polymerase into
RT the Saccharomyces cerevisiae nucleus and specific transcription
RT of target genes";
RL Mol. Cell. Biol. 10:353-360(1990).
XX
RN [21]
RC pJM100 from SIR2 gene
RA Mullen J.R., Kayne P.S., Moerschell R.P., Tsunasawa S., Gribskov M.,
RA Colavito-Shepanski M., Grunstein M., Sherman F., Sternglanz R.;
RT "Identification and characterization of genes and mutants for an
RT N-terminal acetyltransferase from yeast";
RL EMBO J. 8:2067-2075(1989).
XX
RN [22]
RC pJR104 from SIR3 gene
RA Kimmerly W.J., Rine J.;
RT "Replication and segregation of plasmids containing cis-acting
RT regulatory sites of silent mating-type genes in Saccharomyces
RT cerevisiae are controlled by the SIR genes";
RL Mol. Cell. Biol. 7:4225-4237(1987).
XX
RN [23]
RC pJR643 from SIR4 gene
RA Rine J.;
RT ;
RL Unpublished (1991).
XX
RN [24]
RC pSL321 from SNF4 gene
RA Celenza J.L., Eng F.J., Carlson M.;
RT "Molecular analysis of the SNF4 gene of Saccharomyces cerevisiae:
RT evidence for physical association of the SNF4 protein with the
RT SNF1 protein kinase";
RL Mol. Cell. Biol. 9:5045-5054(1989).
XX
RN [25]
RC pCMV-T from CMV
RA Campesi J.;
RT ;
RL Unpublished (1993).
XX
RN [26]
RC pCZ3GAL from CYC1-lacZ gene
RA Chasman D.I., Leatherwood J., Carey M., Ptashne M., Kornberg R.D.;
RT "Activation of yeast polymerase II transcription by herpesvirus VP16
RT and GAL4 derivatives in vitro";
RL Mol. Cell. Biol. 9:4746-4749(1989).
XX
RN [27]
RC pSD.10 from URA3 gene & VP16 domain
RA Dalton S., Treisman R.;
RT "Characterization of SAP-1, a protein recruited by serum response
RT factor to the c-fos serum response element";
RL Cell 68:597-612(1992).
XX
RN [28]
RC pSE1107 from LEU2 gene & GAL4 gene
RA Elledge S.J.;
RT ;
RL Unpublished (1992).
XX
RN [29]
RC pDG95 from HIS3 gene & YEplac195
RA Schiestl R.H., Prakash S.;
RT "RAD1, an excision repair gene of Saccharomyces cerevisiae, is
RT also involved in recombination";
RL Mol. Cell. Biol. 8:3619-3626(1988).
XX
RN [30]
RC pPC62 from LEU2 gene
RC pPC86 from TRP1 gene
RA Chevray P., Nathan D.;
RT "Protein interaction cloning in yeast: identification of mammalian
RT proteins that react with the leucine zipper of Jun";
RL Proc. Natl. Acad. Sci. U.S.A. 89:5789-5793(1992).
XX
RN [31]
RC pACT, pACT2 from LEU2 gene
RA Durfee T., Becherer K., Chen P.L., Yeh S.H., Yang Y., Kilburn A.E.,
RA Lee W.H., Elledge S.J.;
RT "The retinoblastoma protein associates with the protein
RT phosphatase type 1 catalytic subunit";
RL Genes Dev. 7:555-569(1993).
XX
CC NCBI gi: 464015
CC NM (pGAD424)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (CLONTECH)
CC HO (E.coli)
CC CP ()
CC FN (expression)(fusion)
CC SE ()
CC PA ()
CC BR ()
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. yeast, ADH1 promoter SphI-HindIII
FT 2. yeast, GAL4 gene HindIII-EcoRI, activator domain
FT 3. yeast, ADH1 terminator HindIII-SphI
FT 4. pYe(Leu2)10 SalI-XhoI 2200bp, LEU2 gene
FT 5. pBR322, ori/amp
FT 6. yeast 2 micron, ori
FT -> pGAD424 6659bp"
FT misc_binding 1..1
FT /note="SIT SphI"
FT promoter 1..410
FT /note="PRO yeast ADH1 gene"
FT misc_binding 410..410
FT /note="SIT HindIII"
FT promoter 410..830
FT /note="PRO yeast GAL4 ad"
FT misc_binding 480..480
FT /note="SIT unique KpnI"
FT misc_binding 720..720
FT /note="SIT unique MluI"
FT misc_binding 830..858
FT /note="MCS unique EcoRI-SmaI-BamHI-SalI-PstI-BglII"
FT misc_binding 1090..1090
FT /note="SIT HindIII"
FT terminator 1090..1280
FT /note="TER yeast ADH1 gene"
FT misc_binding 1280..1280
FT /note="SIT SphI"
FT CDS 2000..3000
FT /note="YEA yeast LEU2 gene"
FT misc_binding 2220..2220
FT /note="SIT unique EcoRV"
FT misc_binding 2820..2820
FT /note="SIT unique ClaI"
FT misc_binding 3470..3470
FT /note="SIT unique PvuII"
FT rep_origin 3600..3800
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT CDS 4500..5300
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amr)"
FT misc_binding 4660..4660
FT /note="SIT unique BglI"
FT misc_binding 4910..4910
FT /note="SIT unique PvuI"
FT misc_binding 5030..5030
FT /note="SIT unique ScaI"
FT misc_binding 5420..5420
FT /note="SIT unique AatII"
FT rep_origin 0..0
FT /note="ORI yeast 2 micron"
FT misc_binding 6020..6020
FT /note="SIT unique SnaBI"
XX
SQ Sequence 6659 BP; 1902 A; 1499 C; 1367 G; 1891 T; 0 other;
gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga
cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa
gatggataaa gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg
taccgccgcc aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt
cactaacagt agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc
acaaccaatt gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag
taaaattgat gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc
gtataacgcg tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata
taactatcta ttcgatgatg aagatacccc accaaaccca aaaaaagaga tcgaattccc
ggggatccgt cgacctgcag agatctatga atcgtagata ctgaaaaacc ccgcaagttc
acttcaactg tgcatcgtgc accatctcaa tttctttcat ttatacatcg ttttgccttc
ttttatgtaa ctatactcct ctaagtttca atcttggcca tgtaacctct gatctataga
attttttaaa tgactagaat taatgcccat cttttttttg gacctaaatt cttcatgaaa
atatattacg agggcttatt cagaagcttt ggacttcttc gccagaggtt tggtcaagtc
tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
ttaggtttta aaacgaaaat tcttgttctt gagtaactct ttcctgtagg tcaggttgct
ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgcccgaaat
tcccctaccc tatgaacata ttccattttg taatttcgtg tcgtttctat tatgaatttc
atttataaag tttatgtaca aatatcataa aaaaagagaa tctttttaag caaggatttt
cttaacttct tcggcgacag catcaccgac ttcggtggta ctgttggaac cacctaaatc
accagttctg atacctgcat ccaaaacctt tttaactgca tcttcaatgg ccttaccttc
ttcaggcaag ttcaatgaca atttcaacat cattgcagca gacaagatag tggcgatagg
gtcaacctta ttctttggca aatctggagc agaaccgtgg catggttcgt acaaaccaaa
tgcggtgttc ttgtctggca aagaggccaa ggacgcagat ggcaacaaac ccaaggaacc
tgggataacg gaggcttcat cggagatgat atcaccaaac atgttgctgg tgattataat
accatttagg tgggttgggt tcttaactag gatcatggcg gcagaatcaa tcaattgatg
ttgaaccttc aatgtaggaa attcgttctt gatggtttcc tccacagttt ttctccataa
tcttgaagag gccaaaacat tagctttatc caaggaccaa ataggcaatg gtggctcatg
ttgtagggcc atgaaagcgg ccattcttgt gattctttgc acttctggaa cggtgtattg
ttcactatcc caagcgacac catcaccatc gtcttccttt ctcttaccaa agtaaatacc
tcccactaat tctctgacaa caacgaagtc agtaccttta gcaaattgtg gcttgattgg
agataagtct aaaagagagt cggatgcaaa gttacatggt cttaagttgg cgtacaattg
aagttcttta cggattttta gtaaaccttg ttcaggtcta acactacctg taccccattt
aggaccaccc acagcaccta acaaaacggc atcaaccttc ttggaggctt ccagcgcctc
atctggaagt gggacacctg tagcatcgat agcagcacca ccaattaaat gattttcgaa
atcgaacttg acattggaac gaacatcaga aatagcttta agaaccttaa tggcttcggc
tgtgatttct tgaccaacgt ggtcacctgg caaaacgacg atcttcttag gggcagacat
tagaatggta tatccttgaa atatatatat atattgctga aatgtaaaag gtaagaaaag
ttagaaagta agacgattgc taaccaccta ttggaaaaaa caataggtcc ttaaataata
ttgtcaactt caagtattgt gatgcaagca tttagtcatg aacgcttctc tattctatat
gaaaagccgg ttccggcctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg
tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga atttgattct
gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg gttgctaaga
gattcgaact cttgcatctt acgatacctg agtattccca cagttgggga tctcgactct
agctagagga tcaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
tgagtgaggt aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
ctgtcgtgcc agctggatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
gcatcagagc agattgtact gagagtgcac cataacgcat ttaagcataa acacgcacta
tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa
cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg
ctttgaagtt cctattccga agttcctatt ctctagctag aaagtatagg aacttcagag
cgcttttgaa aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg
actgtaacga gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat
ctctttgcta tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc
cttgaacttg catctaaact cgacctctac attttttatg tttatctcta gtattactct
ttagacaaaa aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat
gtaaacattt cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc
tgaccaatga agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca
tcggtataga atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct
agtaatcagt aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac
caaagtagcc ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc
attatagagc gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg
ctctcgggat gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag
cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
tagcgctctc gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa
aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtt
//