LOCUS       ECOLAC                  7477 bp    DNA     linear   BCT 15-NOV-2023
DEFINITION  E.coli lactose operon with lacI, lacZ, lacY and lacA genes.
ACCESSION   J01636 J01637 K01483 K01793
VERSION     J01636.1
KEYWORDS    acetyltransferase; beta-D-galactosidase; galactosidase; lac operon;
            lac repressor protein; lacA gene; lacI gene; lacY gene; lacZ gene;
            lactose permease; mutagenesis; palindrome; promoter region;
            thiogalactoside acetyltransferase.
SOURCE      Escherichia coli
  ORGANISM  Escherichia coli
            Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
            Enterobacteriaceae; Escherichia.
REFERENCE   1  (bases 1243 to 1266)
  AUTHORS   Gilbert,W. and Maxam,A.
  TITLE     The nucleotide sequence of the lac operator
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 70 (12), 3581-3584 (1973)
   PUBMED   4587255
REFERENCE   2  (bases 1246 to 1308)
  AUTHORS   Maizels,N.M.
  TITLE     The nucleotide sequence of the lactose messenger ribonucleic acid
            transcribed from the UV5 promoter mutant of Escherichia coli
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 70 (12), 3585-3589 (1973)
   PUBMED   4587256
REFERENCE   3  (sites)
  AUTHORS   Gilbert,W., Maizels,N. and Maxam,A.
  TITLE     Sequences of controlling regions of the lactose operon
  JOURNAL   Cold Spring Harb. Symp. Quant. Biol. 38, 845-855 (1974)
   PUBMED   4598642
REFERENCE   4  (sites)
  AUTHORS   Gilbert,W., Gralla,J., Majors,A.J. and Maxam,A.
  TITLE     Lactose operator sequences and the action of lac repressor
  JOURNAL   (in) Sund,H. and Blauer,G. (Eds.);
            PROTEIN-LIGAND INTERACTIONS: 193-207;
            Walter de Gruyter, New York (1975)
REFERENCE   5  (bases 1146 to 1282)
  AUTHORS   Dickson,R.C., Abelson,J., Barnes,W.M. and Reznikoff,W.S.
  TITLE     Genetic regulation: the Lac control region
  JOURNAL   Science 187 (4171), 27-35 (1975)
   PUBMED   1088926
REFERENCE   6  (bases 1227 to 1271)
  AUTHORS   Gilbert,W., Maxam,A. and Mirzabekov,A.
  TITLE     Contacts between the lac repressor and DNA revealed by methylation
  JOURNAL   (in) Kjeldgaard,N.C. and Maaloe,O. (Eds.);
            CONTROL OF RIBOSOME SYNTHESIS: 138-143;
            Academic Press, New York (1976)
REFERENCE   7  (sites)
  AUTHORS   Marians,K.J. and Brooker,J.D.
  TITLE     Structure of the lactose operator
  JOURNAL   Nature 260 (5549), 360-363 (1976)
   PUBMED   768781
REFERENCE   8  (bases 1242 to 1268)
  AUTHORS   Heyneker,H.L., Shine,J., Goodman,H.M., Boyer,H.W., Rosenberg,J.,
            Dickerson,R.E., Narang,S.A., Itakura,K., Lin,S. and Riggs,A.D.
  TITLE     Synthetic lac operator DNA is functional in vivo
  JOURNAL   Nature 263 (5580), 748-752 (1976)
   PUBMED   1069185
REFERENCE   9  (sites)
  AUTHORS   Dickson,R.C., Abelson,J. and Johnson,P.
  TITLE     Nucleotide sequence changes produced by mutations in the lac
            promoter of Escherichia coli
  JOURNAL   J. Mol. Biol. 111 (1), 65-75 (1977)
   PUBMED   323498
REFERENCE   10  (bases 51 to 264)
  AUTHORS   Steege,D.A.
  TITLE     5'-Terminal nucleotide sequence of Escherichia coli lactose
            repressor mRNA: features of translational initiation and
            reinitiation sites
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 74 (10), 4163-4167 (1977)
   PUBMED   337294
REFERENCE   11  (bases 1 to 81)
  AUTHORS   Calos,M.P.
  TITLE     DNA sequence for a low-level promoter of the lac repressor gene and
            an 'up' promoter mutation
  JOURNAL   Nature 274 (5673), 762-765 (1978)
   PUBMED   355890
REFERENCE   12  (bases 49 to 1161)
  AUTHORS   Farabaugh,P.J.
  TITLE     Sequence of the lacI gene
  JOURNAL   Nature 274 (5673), 765-769 (1978)
   PUBMED   355891
REFERENCE   13  (sites)
  AUTHORS   Miller,J.H., Coulondre,C. and Farabaugh,P.J.
  TITLE     Correlation of nonsense sites in the lacI gene with specific codons
            in the nucleotide sequence
  JOURNAL   Nature 274 (5673), 770-775 (1978)
   PUBMED   355892
REFERENCE   14  (sites)
  AUTHORS   Calos,M.P. and Miller,J.H.
  TITLE     DNA sequence alteration resulting from a mutation impairing
            promoter function in the lac repressor gene
  JOURNAL   Mol. Gen. Genet. 178 (1), 225-227 (1980)
   PUBMED   6770231
REFERENCE   15  (bases 4306 to 5804)
  AUTHORS   Buchel,D.E., Gronenborn,B. and Muller-Hill,B.
  TITLE     Sequence of the lactose permease gene
  JOURNAL   Nature 283 (5747), 541-545 (1980)
   PUBMED   6444453
REFERENCE   16  (sites)
  AUTHORS   Miller,J.H., Calos,M.P. and Galas,D.J.
  TITLE     Genetic and sequencing studies of the specificity of transposition
            into the lac region of E. coli
  JOURNAL   Cold Spring Harb. Symp. Quant. Biol. 45 (PT 1), 243-257 (1981)
   PUBMED   6271472
REFERENCE   17  (sites)
  AUTHORS   Chenchick,A., Beabealashvilli,R. and Mirzabekov,A.
  TITLE     Topography of interaction of Escherichia coli RNA polymerase
            subunits with lac UV5 promoter
  JOURNAL   FEBS Lett. 128 (1), 46-50 (1981)
   PUBMED   7023981
REFERENCE   18  (sites)
  AUTHORS   Betz,J.L. and Sadler,J.R.
  TITLE     Variants of a cloned synthetic lactose operator. I. A palindromic
            dimer lactose operator derived from one stand of the cloned 40-base
            pair operator
  JOURNAL   Gene 13 (1), 1-12 (1981)
   PUBMED   7016667
REFERENCE   19  (sites)
  AUTHORS   Sadler,J.R. and Tecklenburg,M.
  TITLE     Cloning and characterization of the natural lactose operator
  JOURNAL   Gene 13 (1), 13-23 (1981)
   PUBMED   6263752
REFERENCE   20  (sites)
  AUTHORS   Betz,J.L. and Sadler,J.R.
  TITLE     Variants of a cloned synthetic lactose operator. II.
            Chloramphenicol-resistant revertants retaining a lactose operator
            in the CAT gene of plasmid pBR325
  JOURNAL   Gene 15 (2-3), 187-200 (1981)
   PUBMED   6271642
REFERENCE   21  (sites)
  AUTHORS   Calos,M.P. and Miller,J.H.
  TITLE     The DNA sequence change resulting from the IQ1 mutation, which
            greatly increases promoter strength
  JOURNAL   Mol. Gen. Genet. 183 (3), 559-560 (1981)
   PUBMED   7038381
REFERENCE   22  (sites)
  AUTHORS   Mieschendahl,M., Buchel,D., Bocklage,H. and Muller-Hill,B.
  TITLE     Mutations in the lacY gene of Escherichia coli define functional
            organization of lactose permease
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 78 (12), 7652-7656 (1981)
   PUBMED   6278484
REFERENCE   23  (sites)
  AUTHORS   Russell,D.R. and Bennett,G.N.
  TITLE     Construction and analysis of in vivo activity of E. coli promoter
            hybrids and promoter mutants that alter the -35 to -10 spacing
  JOURNAL   Gene 20 (2), 231-243 (1982)
   PUBMED   6299890
REFERENCE   24  (sites)
  AUTHORS   Horowitz,H. and Platt,T.
  TITLE     A termination site for LacI transcription is between the CAP site
            and the lac promoter
  JOURNAL   J. Biol. Chem. 257 (19), 11740-11746 (1982)
   PUBMED   6288696
REFERENCE   25  (sites)
  AUTHORS   Klein,R.D. and Wells,R.D.
  TITLE     Effects of neighboring DNA homopolymers on the biochemical and
            physical properties of the Escherichia coli lactose promoter. I.
            Cloning and characterization studies
  JOURNAL   J. Biol. Chem. 257 (21), 12954-12961 (1982)
   PUBMED   6290487
REFERENCE   26  (bases 1183 to 1291)
  AUTHORS   Weiher,H. and Schaller,H.
  TITLE     Segment-specific mutagenesis: extensive mutagenesis of a lac
            promoter/operator element
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 79 (5), 1408-1412 (1982)
   PUBMED   7041119
REFERENCE   27  (sites)
  AUTHORS   Van Dyke,M.W. and Dervan,P.B.
  TITLE     Footprinting with MPE.Fe(II). Complementary-strand analyses of
            distamycin- and actinomycin-binding sites on heterogeneous DNA
  JOURNAL   Cold Spring Harb. Symp. Quant. Biol. 47 (PT 1), 347-353 (1983)
   PUBMED   6305557
REFERENCE   28  (bases 1287 to 4364)
  AUTHORS   Kalnins,A., Otto,K., Ruther,U. and Muller-Hill,B.
  TITLE     Sequence of the lacZ gene of Escherichia coli
  JOURNAL   EMBO J. 2 (4), 593-597 (1983)
   PUBMED   6313347
REFERENCE   29  (sites)
  AUTHORS   Cone,K.C., Sellitti,M.A. and Steege,D.A.
  TITLE     Lac repressor mRNA transcription terminates in vivo in the lac
            control region
  JOURNAL   J. Biol. Chem. 258 (18), 11296-11304 (1983)
   PUBMED   6309841
REFERENCE   30  (sites)
  AUTHORS   Sadler,J.R., Sasmor,H. and Betz,J.L.
  TITLE     A perfectly symmetric lac operator binds the lac repressor very
            tightly
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6785-6789 (1983)
   PUBMED   6316325
REFERENCE   31  (sites)
  AUTHORS   Glickman,B.W. and Ripley,L.S.
  TITLE     Structural intermediates of deletion mutagenesis: a role for
            palindromic DNA
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 81 (2), 512-516 (1984)
   PUBMED   6582506
REFERENCE   32  (sites)
  AUTHORS   Spassky,A., Kirkegaard,K. and Buc,H.
  TITLE     Changes in the DNA structure of the lac UV5 promoter during
            formation of an open complex with Escherichia coli RNA polymerase
  JOURNAL   Biochemistry 24 (11), 2723-2731 (1985)
   PUBMED   3896305
REFERENCE   33  (sites)
  AUTHORS   Straney,D.C. and Crothers,D.M.
  TITLE     Intermediates in transcription initiation from the E. coli lac UV5
            promoter
  JOURNAL   Cell 43 (2 PT 1), 449-459 (1985)
   PUBMED   2416465
REFERENCE   34  (sites)
  AUTHORS   Looman,A.C., de Gruyter,M., Vogelaar,A. and van Knippenberg,P.H.
  TITLE     Effects of heterologous ribosomal binding sites on the
            transcription and translation of the lacZ gene of Escherichia coli
  JOURNAL   Gene 37 (1-3), 145-154 (1985)
   PUBMED   3932130
REFERENCE   35  (sites)
  AUTHORS   Mandecki,W., Goldman,R.A., Powell,B.S. and Caruthers,M.H.
  TITLE     lac Up-promoter mutants with increased homology to the consensus
            promoter sequence
  JOURNAL   J. Bacteriol. 164 (3), 1353-1355 (1985)
   PUBMED   2999082
REFERENCE   36  (sites)
  AUTHORS   Malamy,M.H., Rahaim,P.T., Hoffman,C.S., Baghdoyan,D., O'Connor,M.B.
            and Miller,J.F.
  TITLE     A frameshift mutation at the junction of an IS1 insertion within
            lacZ restores beta-galactosidase activity via formation of an
            active lacZ-IS1 fusion protein
  JOURNAL   J. Mol. Biol. 181 (4), 551-555 (1985)
   PUBMED   2987506
REFERENCE   37  (bases 5646 to 7477)
  AUTHORS   Hediger,M.A., Johnson,D.F., Nierlich,D.P. and Zabin,I.
  TITLE     DNA sequence of the lactose operon: the lacA gene and the
            transcriptional termination region
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 82 (19), 6414-6418 (1985)
   PUBMED   3901000
COMMENT     Original source text: Escherichia coli DNA; mRNA; clone
            lambda-h80dlac DNA; clone puk217; pgm8 (see comment).
            [3]  sites; UV5 mRNA transcripts and operator mutants. [(in)
            Sund,H. and Blauer,G. (eds.);Protein-Ligand Interactions:
            193-207;Walter de]  sites; operator mutational analysis. [7]
            sites; S1 and mung bean nuclease action on operator DNA. [9]
            sites; class I, II and III promoter mutant analysis. [13]  sites;
            lacI mutant analysis.
            [16]  sites; Tn5, Tn9 and Tn10 insertion sites in lac region. [14]
            sites; lacI promoter mutation UJ177.
            [18]  sites; palindromic dimer operator;.
            [19]  sites; natural operator sequence.
            [20]  sites; operator mutational analysis.
            [21]  sites; lacI-Q deletion.
            [17]  sites; RNA polymerase UV5 promoter interaction. [22]  sites;
            lacY mutational analysis.
            [24]  sites; lacI-promoted transcription termination. [25]  sites;
            wt and UV5 promoter sequence studies. [23]  sites; UV5 promoter
            mutational analysis.
            [30]  sites; perfectly symmetric operator sequence. [29]  sites;
            lacI mRNA termination site.
            [27]  sites; distamycin and actinomycin binding to promoter. [31]
            sites; lacI deletion studies.
            [35]  sites; promoter mutational studies.
            [33]  sites; DNAase I studies with promoter sequence. [34]  sites;
            ribosomal binding and translation initiation for lacZ. [36]  sites;
            insertion sequence IS1 integration in lacZ;. [32]  sites; DNAase I
            studies with promoter.
            [1] first reports a 27 bp operator(sites 1240-1266) with two-fold
            symmetries; the operator has also been defined to be bases
            1246-1266 or bases 1239-1273 [8]. [(in) Kjeldgaard,N.C. and Maaloe,
            O.(eds);Control of ribosome synthesis: 138-143;A] explores the
            ability of lac
            repressor protein to affect methylation of operator DNA.  [8]
            argues that DNA on both sides of the 21 bp operator (bases
            1246-1266) affects repressor binding but that the sequences of this
            DNA are probably not critical. [5] gives a larger sequence known as
            the promoter-operator region for the wild-type, whereas [2] and
            [26] give portions of this region for the mutant strain UV5. Within
            the promoter region, bases 1162-1199 are identified as the
            catabolite gene activator protein binding site (cap) and bases
            1200-1245 are the RNA polymerase interaction site. [10] reports a
            sequence for the 5'end of the lacI (repressor) gene and discusses
            restart in mutant strains. [11] presents a sequence for the lacI
            promoter region and identifies an I-Q mutation which enhances lacI
            transcription approximately ten-fold. [12] gives a complete
            sequence
            for lacI which agrees with the known lac repressor sequence. [26]
            examines the promoter-operator region in the UV5 strain (lac109)
            and studies 23 mutant derivatives of this sequence. This sequence
            agrees with known protein sequences for the lacZ, lacY and lacA
            enzymes. [15] notes that the fMet codon is not present
            for lacA and suggests that the 'ttg' codon (5727-5729), which
            immediately precedes the mature N-terminal asparagine codon, is the
            start codon. The cds for lacZ, lacY and lacA are included on a
            single mRNA transcript.
            Complete source information:
            Escherichia coli DNA [1],[(in) Kjeldgaard,N.C. and Maaloe,O.(eds);
            Control of ribosome synthesis: 138-143;A],[8],[12],[26]; mRNA [2],
            [5],[10]; clone
            lambda-h80dlac DNA [11],[15]; clone puk217 [28]; pgm8 [37].
FEATURES             Location/Qualifiers
     source          1..7477
                     /organism="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:562"
     variation       16
                     /note="c in wild-type; t in 'up' promoter mutant I-Q [11]"
     gene            79..1161
                     /gene="lacI"
     CDS             79..1161
                     /gene="lacI"
                     /note="lac repressor protein (gtg start codon)"
                     /codon_start=1
                     /transl_table=11
                     /protein_id="AAA24052.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
     regulatory      1162..1199
                     /regulatory_class="other"
                     /note="cap protein binding site"
     variation       1183..1186
                     /note="ttag in wild-type; aatt in strain UV5 [26]"
     variation       1209..1211
                     /note="gct in wild-type; gt in mutant l305 [5]"
     variation       1212
                     /note="t in wild-type; a in mutant l241 [5]"
     variation       1230
                     /note="c in wild-type; a in mutant p-r-1a [5]"
     variation       1237..1238
                     /note="gt in wild-type; aa in strain UV5 [26]"
     variation       1242..1245
                     /note="gtgg in wild-type; ttca in synthetic operator [8]"
     regulatory      1246..1266
                     /regulatory_class="other"
                     /note="lac repressor protein binding site"
     variation       1267..1268
                     /note="tc in wild-type; tg in synthetic operator [8]"
     variation       1282..1291
                     /note="ctatgaccat in wild-type; gatccggcca in strain UV5
                     [26]"
     gene            1284..4358
                     /gene="lacZ"
     CDS             1284..4358
                     /gene="lacZ"
                     /note="beta-d-galactosidase"
                     /codon_start=1
                     /transl_table=11
                     /protein_id="AAA24053.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYT
                     NVTYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWV
                     GYGQDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHK
                     PTTQISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFG
                     GEIIDERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFR
                     EVRIENGLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSH
                     YPNHPLWYTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRN
                     HPSVIIWSLGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVD
                     EDQPFPAVPKWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGG
                     FVWDWVDQSLIKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQ
                     QQFFQFRLSGQTIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIE
                     LPELPQPESAGQLWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPH
                     LTTSEMDFCIELGNKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSE
                     ATRIDPNAWVERWKAAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKT
                     YRIDGSGQMAITVDVEVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAA
                     CFDRWDLPLSDMYTPYVFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETS
                     HRHLLHAEEGTWLNIDGFHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            4410..5663
                     /gene="lacY"
     CDS             4410..5663
                     /gene="lacY"
                     /note="lactose permease"
                     /codon_start=1
                     /transl_table=11
                     /protein_id="AAA24054.1"
                     /translation="MYYLKNTNFWMFGLFFFFYFFIMGAYFPFFPIWLHDINHISKSD
                     TGIIFAAISLFSLLFQPLFGLLSDKLGLRKYLLWIITGMLVMFAPFFIFIFGPLLQYN
                     ILVGSIVGGIYLGFCFNAGAPAVEAFIEKVSRRSNFEFGRARMFGCVGWALCASIVGI
                     MFTINNQFVFWLGSGCALILAVLLFFAKTDAPSSATVANAVGANHSAFSLKLALELFR
                     QPKLWFLSLYVIGVSCTYDVFDQQFANFFTSFFATGEQGTRVFGYVTTMGELLNASIM
                     FFAPLIINRIGGKNALLLAGTIMSVRIIGSSFATSALEVVILKTLHMFEVPFLLVGCF
                     KYITSQFEVRFSATIYLVCFCFFKQLAMIFMSVLAGNMYESIGFQGAYLVLGLVALGF
                     TLISVFTLSGPGPLSLLRRQVNEVA"
     gene            5727..6338
                     /gene="lacA"
     CDS             5727..6338
                     /gene="lacA"
                     /note="thiogalactoside acetyltransferase (ttg start
                     codon)"
                     /codon_start=1
                     /transl_table=11
                     /protein_id="AAA24055.1"
                     /translation="MNMPMTERIRAGKLFTDMCEGLPEKRLRGKTLMYEFNHSHPSEV
                     EKRESLIKEMFATVGENAWVEPPVYFSYGSNIHIGRNFYANFNLTIVDDYTVTIGDNV
                     LIAPNVTLSVTGHPVHHELRKNGEMYSFPITIGNNVWIGSHVVINPGVTIGDNSVIGA
                     GSIVTKDIPPNVVAAGVPCRVIREINDRDKHYYFKDYKVESSV"
BASE COUNT         1739 a         1991 c         2004 g         1743 t
ORIGIN      HindII site [Nature 274, 762-765 (1978)].
        1 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     1201 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1261 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
     1321 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1381 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1441 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
     1501 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1561 tgcacggtta cgatgcgccc atctacacca acgtaaccta tcccattacg gtcaatccgc
     1621 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1681 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttaactcg gcgtttcatc
     1741 tgtggtgcaa cgggcgctgg gtcggttacg gccaggacag tcgtttgccg tctgaatttg
     1801 acctgagcgc atttttacgc gccggagaaa accgcctcgc ggtgatggtg ctgcgttgga
     1861 gtgacggcag ttatctggaa gatcaggata tgtggcggat gagcggcatt ttccgtgacg
     1921 tctcgttgct gcataaaccg actacacaaa tcagcgattt ccatgttgcc actcgcttta
     1981 atgatgattt cagccgcgct gtactggagg ctgaagttca gatgtgcggc gagttgcgtg
     2041 actacctacg ggtaacagtt tctttatggc agggtgaaac gcaggtcgcc agcggcaccg
     2101 cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta tgccgatcgc gtcacactac
     2161 gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat cccgaatctc tatcgtgcgg
     2221 tggttgaact gcacaccgcc gacggcacgc tgattgaagc agaagcctgc gatgtcggtt
     2281 tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa cggcaagccg ttgctgattc
     2341 gaggcgttaa ccgtcacgag catcatcctc tgcatggtca ggtcatggat gagcagacga
     2401 tggtgcagga tatcctgctg atgaagcaga acaactttaa cgccgtgcgc tgttcgcatt
     2461 atccgaacca tccgctgtgg tacacgctgt gcgaccgcta cggcctgtat gtggtggatg
     2521 aagccaatat tgaaacccac ggcatggtgc caatgaatcg tctgaccgat gatccgcgct
     2581 ggctaccggc gatgagcgaa cgcgtaacgc gaatggtgca gcgcgatcgt aatcacccga
     2641 gtgtgatcat ctggtcgctg gggaatgaat caggccacgg cgctaatcac gacgcgctgt
     2701 atcgctggat caaatctgtc gatccttccc gcccggtgca gtatgaaggc ggcggagccg
     2761 acaccacggc caccgatatt atttgcccga tgtacgcgcg cgtggatgaa gaccagccct
     2821 tcccggctgt gccgaaatgg tccatcaaaa aatggctttc gctacctgga gagacgcgcc
     2881 cgctgatcct ttgcgaatac gcccacgcga tgggtaacag tcttggcggt ttcgctaaat
     2941 actggcaggc gtttcgtcag tatccccgtt tacagggcgg cttcgtctgg gactgggtgg
     3001 atcagtcgct gattaaatat gatgaaaacg gcaacccgtg gtcggcttac ggcggtgatt
     3061 ttggcgatac gccgaacgat cgccagttct gtatgaacgg tctggtcttt gccgaccgca
     3121 cgccgcatcc agcgctgacg gaagcaaaac accagcagca gtttttccag ttccgtttat
     3181 ccgggcaaac catcgaagtg accagcgaat acctgttccg tcatagcgat aacgagctcc
     3241 tgcactggat ggtggcgctg gatggtaagc cgctggcaag cggtgaagtg cctctggatg
     3301 tcgctccaca aggtaaacag ttgattgaac tgcctgaact accgcagccg gagagcgccg
     3361 ggcaactctg gctcacagta cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg
     3421 ggcacatcag cgcctggcag cagtggcgtc tggcggaaaa cctcagtgtg acgctccccg
     3481 ccgcgtccca cgccatcccg catctgacca ccagcgaaat ggatttttgc atcgagctgg
     3541 gtaataagcg ttggcaattt aaccgccagt caggctttct ttcacagatg tggattggcg
     3601 ataaaaaaca actgctgacg ccgctgcgcg atcagttcac ccgtgcaccg ctggataacg
     3661 acattggcgt aagtgaagcg acccgcattg accctaacgc ctgggtcgaa cgctggaagg
     3721 cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat acacttgctg
     3781 atgcggtgct gattacgacc gctcacgcgt ggcagcatca ggggaaaacc ttatttatca
     3841 gccggaaaac ctaccggatt gatggtagtg gtcaaatggc gattaccgtt gatgttgaag
     3901 tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag
     3961 cagagcgggt aaactggctc ggattagggc cgcaagaaaa ctatcccgac cgccttactg
     4021 ccgcctgttt tgaccgctgg gatctgccat tgtcagacat gtataccccg tacgtcttcc
     4081 cgagcgaaaa cggtctgcgc tgcgggacgc gcgaattgaa ttatggccca caccagtggc
     4141 gcggcgactt ccagttcaac atcagccgct acagtcaaca gcaactgatg gaaaccagcc
     4201 atcgccatct gctgcacgcg gaagaaggca catggctgaa tatcgacggt ttccatatgg
     4261 ggattggtgg cgacgactcc tggagcccgt cagtatcggc ggaattccag ctgagcgccg
     4321 gtcgctacca ttaccagttg gtctggtgtc aaaaataata ataaccgggc aggccatgtc
     4381 tgcccgtatt tcgcgtaagg aaatccatta tgtactattt aaaaaacaca aacttttgga
     4441 tgttcggttt attctttttc ttttactttt ttatcatggg agcctacttc ccgtttttcc
     4501 cgatttggct acatgacatc aaccatatca gcaaaagtga tacgggtatt atttttgccg
     4561 ctatttctct gttctcgcta ttattccaac cgctgtttgg tctgctttct gacaaactcg
     4621 ggctgcgcaa atacctgctg tggattatta ccggcatgtt agtgatgttt gcgccgttct
     4681 ttatttttat cttcgggcca ctgttacaat acaacatttt agtaggatcg attgttggtg
     4741 gtatttatct aggcttttgt tttaacgccg gtgcgccagc agtagaggca tttattgaga
     4801 aagtcagccg tcgcagtaat ttcgaatttg gtcgcgcgcg gatgtttggc tgtgttggct
     4861 gggcgctgtg tgcctcgatt gtcggcatca tgttcaccat caataatcag tttgttttct
     4921 ggctgggctc tggctgtgca ctcatcctcg ccgttttact ctttttcgcc aaaacggatg
     4981 cgccctcttc tgccacggtt gccaatgcgg taggtgccaa ccattcggca tttagcctta
     5041 agctggcact ggaactgttc agacagccaa aactgtggtt tttgtcactg tatgttattg
     5101 gcgtttcctg cacctacgat gtttttgacc aacagtttgc taatttcttt acttcgttct
     5161 ttgctaccgg tgaacagggt acgcgggtat ttggctacgt aacgacaatg ggcgaattac
     5221 ttaacgcctc gattatgttc tttgcgccac tgatcattaa tcgcatcggt gggaaaaacg
     5281 ccctgctgct ggctggcact attatgtctg tacgtattat tggctcatcg ttcgccacct
     5341 cagcgctgga agtggttatt ctgaaaacgc tgcatatgtt tgaagtaccg ttcctgctgg
     5401 tgggctgctt taaatatatt accagccagt ttgaagtgcg tttttcagcg acgatttatc
     5461 tggtctgttt ctgcttcttt aagcaactgg cgatgatttt tatgtctgta ctggcgggca
     5521 atatgtatga aagcatcggt ttccagggcg cttatctggt gctgggtctg gtggcgctgg
     5581 gcttcacctt aatttccgtg ttcacgctta gcggccccgg cccgctttcc ctgctgcgtc
     5641 gtcaggtgaa tgaagtcgct taagcaatca atgtcggatg cggcgcgacg cttatccgac
     5701 caacatatca taacggagtg atcgcattga acatgccaat gaccgaaaga ataagagcag
     5761 gcaagctatt taccgatatg tgcgaaggct taccggaaaa aagacttcgt gggaaaacgt
     5821 taatgtatga gtttaatcac tcgcatccat cagaagttga aaaaagagaa agcctgatta
     5881 aagaaatgtt tgccacggta ggggaaaacg cctgggtaga accgcctgtc tatttctctt
     5941 acggttccaa catccatata ggccgcaatt tttatgcaaa tttcaattta accattgtcg
     6001 atgactacac ggtaacaatc ggtgataacg tactgattgc acccaacgtt actctttccg
     6061 ttacgggaca ccctgtacac catgaattga gaaaaaacgg cgagatgtac tcttttccga
     6121 taacgattgg caataacgtc tggatcggaa gtcatgtggt tattaatcca ggcgtcacca
     6181 tcggggataa ttctgttatt ggcgcgggta gtatcgtcac aaaagacatt ccaccaaacg
     6241 tcgtggcggc tggcgttcct tgtcgggtta ttcgcgaaat aaacgaccgg gataagcact
     6301 attatttcaa agattataaa gttgaatcgt cagtttaaat tataaaaatt gcctgatacg
     6361 ctgcgcttat caggcctaca agttcagcga tctacattag ccgcatccgg catgaacaaa
     6421 gcgcaggaac aagcgtcgca tcatgcctct ttgacccaca gctgcggaaa acgtactggt
     6481 gcaaaacgca gggttatgat catcagccca acgacgcaca gcgcatgaaa tgcccagtcc
     6541 atcaggtaat tgccgctgat actacgcagc acgccagaaa accacggggc aagcccggcg
     6601 atgataaaac cgattccctg cataaacgcc accagcttgc cagcaatagc cggttgcaca
     6661 gagtgatcga gcgccagcag caaacagagc ggaaacgcgc cgcccagacc taacccacac
     6721 accatcgccc acaataccgg caattgcatc ggcagccaga taaagccgca gaaccccacc
     6781 agttgtaaca ccagcgccag cattaacagt ttgcgccgat cctgatggcg agccatagca
     6841 ggcatcagca aagctcctgc ggcttgccca agcgtcatca atgccagtaa ggaaccgctg
     6901 tactgcgcgc tggcaccaat ctcaatatag aaagcgggta accaggcaat caggctggcg
     6961 taaccgccgt taatcagacc gaagtaaaca cccagcgtcc acgcgcgggg agtgaatacc
     7021 acgcgaaccg gagtggttgt tgtcttgtgg gaagaggcga cctcgcgggc gctttgccac
     7081 caccaggcaa agagcgcaac aacggcaggc agcgccacca ggcgagtgtt tgataccagg
     7141 tttcgctatg ttgaactaac cagggcgtta tggcggcacc aagcccaccg ccgcccatca
     7201 gagccgcgga ccacagcccc atcaccagtg gcgtgcgctg ctgaaaccgc cgtttaatca
     7261 ccgaagcatc accgcctgaa tgatgccgat ccccacccca ccaagcagtg cgctgctaag
     7321 cagcagcgca ctttgcgggt aaagctcacg catcaatgca ccgacggcaa tcagcaacag
     7381 actgatggcg acactgcgac gttcgctgac atgctgatga agccagcttc cggccagcgc
     7441 cagcccgccc atggtaacca ccggcagagc ggtcgac
//