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ID   PGAD424    preliminary; circular DNA; SYN; 6659 BP.
AC   U07647;
DT   01-JUL-1994 (Rel. 9, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Yeast/E.coli plasmid vector pGAD424 - complete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RL   Unpublished (1994).
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.
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.
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).
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).
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).
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).
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).
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).
RN   [10]
RC   [from none]
RA   Fritz C.C., Green M.R.;
RT   ;
RL   Curr. Biol. 2:403-405(1992).
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).
RN   [12]
RC   pRY121-2micron- from pRY121
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).
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).
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).
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).
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).
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).
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).
RN   [19]
RC   pES15 from pUC18 & SIR1 gene
RA   Stone E.;
RT   ;
RL   Unpublished (1991).
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).
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).
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).
RN   [23]
RC   pJR643 from SIR4 gene
RA   Rine J.;
RT   ;
RL   Unpublished (1991).
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).
RN   [25]
RC   pCMV-T from CMV
RA   Campesi J.;
RT   ;
RL   Unpublished (1993).
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).
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).
RN   [28]
RC   pSE1107 from LEU2 gene & GAL4 gene
RA   Elledge S.J.;
RT   ;
RL   Unpublished (1992).
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).
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).
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).
CC   NCBI gi: 464015
CC   NM (pGAD424)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli)
CC   CP ()
CC   FN (expression)(fusion)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
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"
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