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LOCUS SYNPBR322 4361 bp DNA circular SYN 30-SEP-2008 DEFINITION Cloning vector pBR322, complete sequence. ACCESSION J01749 K00005 L08654 M10282 M10283 M10286 M10356 M10784 M10785 M10786 M33694 V01119 VERSION J01749.1 KEYWORDS ampicillin resistance; beta-lactamase; cloning vector; drug resistance protein; origin of replication; plasmid; tetracycline resistance. SOURCE Cloning vector pBR322 ORGANISM Cloning vector pBR322 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 3; 3259 to 4361) AUTHORS Sutcliffe,J.G. TITLE Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322 JOURNAL Proc. Natl. Acad. Sci. U.S.A. 75 (8), 3737-3741 (1978) PUBMED 358200 REFERENCE 2 (bases 1 to 4361) AUTHORS Sutcliffe,J.G. TITLE Complete nucleotide sequence of the Escherichia coli plasmid pBR322 JOURNAL Cold Spring Harb. Symp. Quant. Biol. 43 (Pt 1), 77-90 (1979) PUBMED 383387 REFERENCE 3 (bases 1500 to 2300) AUTHORS Reed,R.R., Young,R.A., Steitz,J.A., Grindley,N.D. and Guyer,M.S. TITLE Transposition of the Escherichia coli insertion element gamma generates a five-base-pair repeat JOURNAL Proc. Natl. Acad. Sci. U.S.A. 76 (10), 4882-4886 (1979) PUBMED 388421 REFERENCE 4 (bases 2207 to 2265) AUTHORS Covarrubias,L., Cervantes,L., Covarrubias,A., Soberon,X., Vichido,I., Blanco,A., Kupersztoch-Portnoy,Y.M. and Bolivar,F. TITLE Construction and characterization of new cloning vehicles. V. Mobilization and coding properties of pBR322 and several deletion derivatives including pBR327 and pBR328 JOURNAL Gene 13 (1), 25-35 (1981) PUBMED 6263753 REFERENCE 5 (bases 2000 to 2500) AUTHORS Marians,K.J., Soeller,W. and Zipursky,S.L. TITLE Maximal limits of the Escherichia coli replication factor Y effector site sequences in pBR322 DNA JOURNAL J. Biol. Chem. 257 (10), 5656-5662 (1982) PUBMED 6279609 REFERENCE 6 (bases 1 to 80; 4151 to 4229; 4349 to 4361) AUTHORS Brosius,J., Cate,R.L. and Perlmutter,A.P. TITLE Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro JOURNAL J. Biol. Chem. 257 (15), 9205-9210 (1982) PUBMED 6178738 REFERENCE 7 (bases 4241 to 4343) AUTHORS Van Dyke,M.W., Hertzberg,R.P. and Dervan,P.B. TITLE Map of distamycin, netropsin, and actinomycin binding sites on heterogeneous DNA: DNA cleavage-inhibition patterns with methidiumpropyl-EDTA.Fe(II) JOURNAL Proc. Natl. Acad. Sci. U.S.A. 79 (18), 5470-5474 (1982) PUBMED 6291045 REFERENCE 8 (bases 584 to 709) AUTHORS Peden,K.W. and Nathans,D. TITLE Local mutagenesis within deletion loops of DNA heteroduplexes JOURNAL Proc. Natl. Acad. Sci. U.S.A. 79 (23), 7214-7217 (1982) PUBMED 6760191 REFERENCE 9 (bases 373 to 649) AUTHORS Peden,K.W. TITLE Revised sequence of the tetracycline-resistance gene of pBR322 JOURNAL Gene 22 (2-3), 277-280 (1983) PUBMED 6307828 REFERENCE 10 (bases 132 to 181) AUTHORS Watabe,H., Iino,T., Kaneko,T., Shibata,T. and Ando,T. TITLE A new class of site-specific endodeoxyribonucleases. Endo.Sce I isolated from a eukaryote, Saccharomyces cerevisiae JOURNAL J. Biol. Chem. 258 (8), 4663-4665 (1983) PUBMED 6300094 REFERENCE 11 (bases 368 to 581) AUTHORS Livneh,Z. TITLE Directed mutagenesis method for analysis of mutagen specificity: application to ultraviolet-induced mutagenesis JOURNAL Proc. Natl. Acad. Sci. U.S.A. 80 (1), 237-241 (1983) PUBMED 6337373 REFERENCE 12 (bases 2627 to 2682; 2781 to 2828) AUTHORS Mascharak,P.K., Sugiura,Y., Kuwahara,J., Suzuki,T. and Lippard,S.J. TITLE Alteration and activation of sequence-specific cleavage of DNA by bleomycin in the presence of the antitumor drug cis-diamminedichloroplatinum(II) JOURNAL Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6795-6798 (1983) PUBMED 6196777 REFERENCE 13 (bases 4276 to 4336) AUTHORS Schultz,P.G. and Dervan,P.B. TITLE Sequence-specific double-strand cleavage of DNA by penta-N-methylpyrrolecarboxamide-EDTA X Fe(II) JOURNAL Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6834-6837 (1983) PUBMED 6417654 REFERENCE 14 (bases 518 to 528) AUTHORS Sutcliffe,J.G. JOURNAL Unpublished REFERENCE 15 (bases 2395 to 2495) AUTHORS Fuller,R.S., Funnell,B.E. and Kornberg,A. TITLE The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites JOURNAL Cell 38 (3), 889-900 (1984) PUBMED 6091903 REFERENCE 16 (bases 2729 to 2731) AUTHORS Lathe,R., Kieny,M.P., Skory,S. and Lecocq,J.P. TITLE Linker tailing: unphosphorylated linker oligonucleotides for joining DNA termini JOURNAL DNA 3 (2), 173-182 (1984) PUBMED 6327214 REFERENCE 17 (bases 2729 to 2730) AUTHORS Heusterspreute,M. and Davison,J. TITLE Restriction site bank vectors. II. DNA sequence analysis of plasmid pJRD158 JOURNAL DNA 3 (3), 259-268 (1984) PUBMED 6086259 REFERENCE 18 (bases 2113 to 2186; 2348 to 2415) AUTHORS Abarzua,P., Soeller,W. and Marians,K.J. TITLE Mutational analysis of primosome assembly sites. I. Distinct classes of mutants in the pBR322 Escherichia coli factor Y DNA effector sequences JOURNAL J. Biol. Chem. 259 (22), 14286-14292 (1984) PUBMED 6209275 REFERENCE 19 (bases 2348 to 2415) AUTHORS Soeller,W., Abarzua,P. and Marians,K.J. TITLE Mutational analysis of primosome assembly sites. II. Role of secondary structure in the formation of active sites JOURNAL J. Biol. Chem. 259 (22), 14293-14300 (1984) PUBMED 6150042 REFERENCE 20 (bases 1 to 4361) AUTHORS Van Dyke,M.M. and Dervan,P.B. TITLE Echinomycin binding sites on DNA JOURNAL Science 225 (4667), 1122-1127 (1984) PUBMED 6089341 REFERENCE 21 (sites) AUTHORS Pouwels,P.H., Enger-Valk,B.E. and Brammar,W.J. JOURNAL (in) CLONING VECTORS. Elsevier Scientific Publishing, Amsterdam (1985) REMARK Vector I-A-iv-1 REFERENCE 22 (bases 1 to 4361) AUTHORS Watson,N. TITLE A new revision of the sequence of plasmid pBR322 JOURNAL Gene 70 (2), 399-403 (1988) PUBMED 3063608 REFERENCE 23 (sites) AUTHORS Gilbert,W. TITLE Obtained from VecBase 3.0 JOURNAL Unpublished COMMENT On or before Apr 4, 2002 this sequence version replaced L08654.1, gi:58257. The circular sequence is numbered such that 0 is the middle of the unique EcoRI site and the count increases first through the tet genes, the pMB1 material, and finally through the Tn3 region. Plasmid pBR322 contains ampicillin and tetracycline resistance genes. The ampicillin resistance gene (amp-r) is a penicillin beta-lactamase. Promoters P1 and P3 are for the beta-lactamase gene. P3 is the natural promoter, and P1 is artificially created by the ligation of two different DNA fragments to create pBR322. P2 is in the same region as P1, but it is on the opposite strand and initiates transcription in the direction of the tetracycline resistance gene. Mutational studies in the primosome assembly sites indicate four types of mutations: Class I having no effect on the activities elicited by the DNA site and the bases involved are probably spacers; Class II requiring higher Mg-2+ concentrations than the wild-type to be fully activated as factor Y ATPase effectors; Class III co-inactivating both the ATPase effector and DNA replication template activity of the site, indicating that they probably represent essential contact points between factor Y and the DNA; Class IV having a replication template activity intermediate that of class III and class II mutant DNAs. Specific sites within or near the origins of replication are recognized by dnaA protein. Without dnaA binding to the origin of replication chromosomal replication is not possible (15). pBR322 DNA contains two separate regions on opposite strands and close to the origin of replication which, when in single-stranded form, can act as effectors for the ATPase activity of E.coli replication factor Y (5). Small fragments of DNA containing these sites when cloned in an f1 phage vector act as origins of DNA replication allowing the formation of complementary double-stranded DNA in rifampicin-resistant, dna(B,G,C)-dependent fashion in vitro (5). The biological activity of echinomycin is thought to be related to the formation of complexes by intercalating with cellular DNA (20). Complete source information: Plasmid pBR322 from E.coli (2),(1),(3),(6),(11),(8),(5),(7),(12), (13),(10),(9),(14),(18),(19),(15),(20),(16); pBR322 DNA in pXf3 (4). The following data and their annotation were supplied by Will Gilbert under the auspices of the Curator Program. CROSSREFERENCE #parent GenBank(50):pSC101C, GenBank(50):Trn3 #offspring VecBase(3):pBR325, VecBase(3):pBR327, VecBase(3):pBR328, VecBase(3):pAT153, VecBase(3):pUC7, VecBase(3):pJRD158, VecBase(3):PiVX, VecBase(3):PiAN7, VecBase(3):pSP64, VecBase(3):pSP65, VecBase(3):pGEM1, VecBase(3):pGEM2, VecBase(3):pGEM3, VecBase(3):pGEM4, VecBase(3):pKK223, VecBase(3):pLBU3, VecBase(3):pTrS3, VecBase(3):pRSVNeo, VecBase(3):pSV2Cat, VecBase(3):M13mp9, VecBase(3):pHC79, VecBase(3):pV34, VecBase(3):pKTH601, VecBase(3):pKTH604, VecBase(3):pKTH605, VecBase(3):pKTH606, VecBase(3):YEp24, VecBase(3):YIp5, VecBase(3):YRp17, VecBase(3):pSP18, VecBase(3):pSP19, VecBase(3):pSP6T3, VecBase(3):pSP6T719, VecBase(3):pT712, VecBase(3):pT713, VecBase(3):pT7T318, VecBase(3):pT7T319, VecBase(3):pT7T3A18, VecBase(3):pT7T3A19, VecBase(3):pEX1, VecBase(3):pEX2, VecBase(3):pEX3, VecBase(3):pCKSP6, VecBase(3):pACYC177, VecBase(3):pKO1, VecBase(3):pKO2, VecBase(3):pKM1, VecBase(3):pKM2, VecBase(3):pMBL1, VecBase(3):pMBL604, VecBase(3):pMC1511, VecBase(3):pMC1871, VecBase(3):pAA37X, VecBase(3):pUR278, VecBase(3):pUR288, VecBase(3):pUR289, VecBase(3):pUR290, VecBase(3):pUR291, VecBase(3):pUR292, VecBase(3):pUR222. FEATURES Location/Qualifiers source 1..4361 /organism="Cloning vector pBR322" /mol_type="other DNA" /db_xref="taxon:47470" /tissue_lib="ATCC 31344, ATCC 37017" /focus source 1..1762 /organism="Plasmid pSC101" /mol_type="other DNA" /db_xref="taxon:2625" misc_binding 24..27 /bound_moiety="echinomycin" regulatory complement(27..33) /regulatory_class="promoter" /note="promoter P1 (6)" misc_binding 39..42 /bound_moiety="echinomycin" regulatory 43..49 /regulatory_class="promoter" /note="promoter P2 (6)" misc_binding 53..56 /bound_moiety="echinomycin" misc_binding 67..70 /bound_moiety="echinomycin" misc_binding 80..83 /bound_moiety="echinomycin" gene 86..1276 /gene="tet" CDS 86..1276 /gene="tet" /codon_start=1 /transl_table=11 /product="tetracycline resistance protein" /protein_id="AAB59735.1" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG QLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR ATST" misc_feature complement(141..142) /note="Endo.Sce I cleavage site coordinated with site at base 146 (10)" misc_feature 146..147 /gene="tet" /note="Endo.Sce I cleavage site coordinated with site at base 142 (10)" misc_binding 411..414 /gene="tet" /bound_moiety="echinomycin" misc_difference 426 /gene="tet" /note="conflict" /citation=[11] /replace="" misc_binding 469..472 /gene="tet" /bound_moiety="echinomycin" old_sequence 526..528 /gene="tet" /citation=[17] repeat_region complement(1515..1519) /note="gamma-delta insertion target sequence" /rpt_type=direct misc_feature 1636..1762 /note="from pSC101 (bp 1860-1986)" repeat_region complement(1788..1792) /note="gamma-delta insertion target sequence" /rpt_type=direct misc_difference 1891..1892 /note="conflict" /citation=[23] /replace="att" old_sequence 1892..1893 /citation=[2] /citation=[22] regulatory 1905..1910 /regulatory_class="ribosome_binding_site" regulatory 1905..1909 /regulatory_class="ribosome_binding_site" /note="Shine-Dalgarno sequence" misc_difference 1913..1914 /note="conflict" /citation=[23] /replace="caa" old_sequence 1914..1915 /citation=[17] CDS 1915..2106 /codon_start=1 /transl_table=11 /product="ROP protein" /protein_id="AAB59736.1" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" misc_feature 2011..2167 /note="H-strand Y effector site" /citation=[5] repeat_region complement(2245..2249) /note="gamma-delta insertion target sequence" /rpt_type=direct misc_feature complement(2351..2414) /note="L-strand Y effector site" /citation=[5] misc_binding 2439..2447 /bound_moiety="dnaA" rep_origin 2535 old_sequence 2729..2730 /note="revision according to (17)" /citation=[2] /citation=[17] /replace="at" old_sequence 2729 /citation=[17] old_sequence 2730 /note="revision according to (16)" /citation=[2] /citation=[16] /citation=[17] /replace="t" mobile_element 3148..4361 /mobile_element_type="transposon:Tn3" repeat_region 3148..3185 /note="corresponds to one of the 38bp repeats found in Tn3 (bp 1-38 and complement (4920-4957))" /rpt_type=inverted gene complement(3293..4153) /gene="bla" CDS complement(3293..4153) /gene="bla" /note="E-286" /codon_start=1 /transl_table=11 /product="beta-lactamase" /protein_id="AAB59737.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide complement(4085..4153) /gene="bla" mat_peptide complement(3296..4084) /gene="bla" /product="beta-lactamase" regulatory complement(4161..4165) /regulatory_class="ribosome_binding_site" /note="Shine-Dalgarno sequence" regulatory complement(4188..4194) /regulatory_class="promoter" /note="promoter P3 (6)" misc_binding complement(4268..4271) /bound_moiety="echinomycin" misc_binding complement(4280..4283) /bound_moiety="echinomycin" misc_binding complement(4285..4288) /bound_moiety="echinomycin" misc_binding complement(4296..4299) /bound_moiety="echinomycin" misc_binding complement(4311..4314) /bound_moiety="echinomycin" misc_binding complement(4317..4320) /bound_moiety="echinomycin" misc_binding complement(4331..4334) /bound_moiety="echinomycin" ORIGIN 1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa 61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg 121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt 181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata 241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg 301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac 361 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac 421 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca 481 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg 541 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct 601 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat 661 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt 721 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct 781 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct 841 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa 901 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt 961 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc 1021 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca 1081 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc 1141 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat 1201 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg 1261 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga 1321 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac 1381 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg 1441 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg 1501 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc 1561 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg 1621 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca 1681 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga 1741 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa 1801 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt 1861 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc 1921 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt 1981 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac 2041 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac 2101 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 2161 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc 2221 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg 2281 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 2341 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 2401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 2581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 2641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 2701 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 2761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 2821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 2881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 2941 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 3001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 3241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 3301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 3361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 3421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 3481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 3541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 3601 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 3661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 3721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 3781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 3841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 3901 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 3961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 4021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 4081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 4141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 4201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 4261 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 4321 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga a //