LOCUS ECOLAC 7477 bp DNA linear BCT 05-MAY-1993 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; Proteobacteria; 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]" mRNA 51..1230 /note="lacI (repressor) mRNA; preferred in vivo 3' end [12],[29]" 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]" mRNA 1246..>4358 /note="lacZ mRNA [2],[5]" 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" 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 //