File: CVPBR322

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ID   CVPBR322   standard; circular DNA; SYN; 4361 BP.
XX
AC   J01749; K00005; L08654; M10282; M10283; M10286; M10356; M10784;
AC   M10785; M10786; M33694; V01119;
XX
DT   16-JUN-1990 (Rel. 24, Created)
DT   02-AUG-1995 (Rel. 44, Last updated, Version 20)
XX
DE   Cloning vector pBR322, complete genome.
XX
KW   ampicillin resistance; beta-lactamase; cloning vector;
KW   drug resistance protein; origin of replication; plasmid;
KW   tetracycline resistance.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vectors.
XX
RN   [1]
RP   1-3, 3259-4361
RX   MEDLINE; 79012484.
RA   Sutcliffe J.G.;
RT   "Nucleotide sequence of the ampicillin resistance gene of
RT   Escherichia coli plasmid pBR322";
RL   Proc. Natl. Acad. Sci. U.S.A. 75:3737-3741(1978).
XX
RN   [2]
RP   1-4361
RX   MEDLINE; 80002802.
RA   Sutcliffe J.G.;
RT   "Complete nucleotide sequence of the Escherichia coli plasmid
RT   pBR322";
RL   Cold Spring Harb. Symp. Quant. Biol. 43:77-90(1979).
XX
RN   [3]
RP   1500-2300
RX   MEDLINE; 80056597.
RA   Reed R.R., Young R.A., Steitz J.A., Grindley N.D.F., Guyer M.;
RT   "Transposition of the Escherichia coli insertion element gamma
RT   generates a five-base-pair repeat";
RL   Proc. Natl. Acad. Sci. U.S.A. 76:4882-4886(1979).
XX
RN   [4]
RP   2207-2265
RX   MEDLINE; 81213464.
RA   Covarrubias L., Cervantes L., Covarrubias A., Soberon X.,
RA   Vichido I., Blanco A., Kupersztoch-Portnoy Y.M., Bolivar F.;
RT   "Construction and characterization of new cloning vehicles: V.
RT   Mobilization and coding properties of pBR322 and several deletion
RT   derivatives including pBR327 and pBR328";
RL   Gene 13:25-35(1981).
XX
RN   [5]
RP   2000-2500
RX   MEDLINE; 82167416.
RA   Marians K.J., Soeller W., Zipursky S.L.;
RT   "Maximal limits of the Escherichia coli replication factor Y
RT   effector site sequences in pBR322 DNA";
RL   J. Biol. Chem. 257:5656-5662(1982).
XX
RN   [6]
RP   1-80, 4151-4229, 4349-4361
RX   MEDLINE; 82239419.
RA   Brosius J., Cate R.L., Perlmutter A.P.;
RT   "Precise location of two promoters for the beta-lactamase gene of
RT   pBR322 : S1 mapping of ribonucleic acid isolated from Escherichia
RT   coli or synthesized in vitro";
RL   J. Biol. Chem. 257:9205-9210(1982).
XX
RN   [7]
RP   4241-4343
RX   MEDLINE; 83039392.
RA   Van dyke M.W., Hertzberg R.P., Dervan P.B.;
RT   "Map of distamycin, netropsin, and actinomycin binding sites on
RT   heterogeneous DNA: Dna cleavage-inhibition patterns with
RT   methidiumpropyl-EDTA.Fe(II)";
RL   Proc. Natl. Acad. Sci. U.S.A. 79:5470-5474(1982).
XX
RN   [8]
RP   584-709
RX   MEDLINE; 83117649.
RA   Peden K.W.C., Nathans D.;
RT   "Local mutagenesis within deletion loops of DNA heteroduplexes";
RL   Proc. Natl. Acad. Sci. U.S.A. 79:7214-7217(1982).
XX
RN   [9]
RP   373-649
RX   MEDLINE; 83263146.
RA   Peden K.W.C.;
RT   "Revised sequence of the tetracycline-resistance gene of pBR322";
RL   Gene 22:277-280(1983).
XX
RN   [10]
RP   132-181
RX   MEDLINE; 83161053.
RA   Watabe H.o., Iino T., Kaneko T., Shibata T., Ando T.;
RT   "A new class of site-specific endo-deoxyribonucleases: Endo.Sce I
RT   isolated from a eukaryote, Saccharomyces cerevisiae";
RL   J. Biol. Chem. 258:4663-4665(1983).
XX
RN   [11]
RP   368-581
RX   MEDLINE; 83117828.
RA   Livneh Z.;
RT   "Directed mutagenesis method for analysis of mutagen specificity:
RT   Application to ultraviolet-induced mutagenesis";
RL   Proc. Natl. Acad. Sci. U.S.A. 80:237-241(1983).
XX
RN   [12]
RP   2627-2682, 2781-2828
RX   MEDLINE; 84070716.
RA   Mascharak P.K., Sugiura Y., Kuwahara J., Suzuki T., Lippard S.J.;
RT   "Alteration and activation of sequence-specific cleavage of DNA by
RT   bleomycin in the presence of the antitumor drug
RT   cis-diamminedichloroplatinum(II)";
RL   Proc. Natl. Acad. Sci. U.S.A. 80:6795-6798(1983).
XX
RN   [13]
RP   4276-4336
RX   MEDLINE; 84070724.
RA   Schultz P.G., Dervan P.B.;
RT   "Sequence-specific double-stranded cleavage of DNA by
RT   penta-N-methylpyrrolecarboxamide-EDTA.Fe(II)";
RL   Proc. Natl. Acad. Sci. U.S.A. 80:6834-6837(1983).
XX
RN   [14]
RP   518-528
RA   Sutcliffe J.G.;
RT   ;
RL   Unpublished.
XX
RN   [15]
RP   2395-2495
RX   MEDLINE; 85024881.
RA   Fuller R.S., Funnell B.E., Kornberg A.;
RT   "The dnaA protein complex with the E. coli chromosomal replication
RT   origin (oriC) and other DNA sites";
RL   Cell 38:889-900(1984).
XX
RN   [16]
RP   2729-2731
RX   MEDLINE; 84207440.
RA   Lathe R., Kieny M.P., Skory S., Lecocoq J.P.;
RT   "Laboratory methods linker tailing: Unphosphorylated linker
RT   oligonucleotides for joining DNA termini";
RL   DNA 3:173-182(1984).
XX
RN   [17]
RC   bases
RX   MEDLINE; 84260952.
RA   Heusterspreute M., Davison J.;
RT   "Restriction site bank vectors. II. DNA sequence analysis of
RT   plasmid pJRD158";
RL   DNA 3:259-268(1984).
XX
RN   [18]
RP   2113-2186, 2348-2415
RX   MEDLINE; 85054885.
RA   Abarzua P.I., Soeller W., Marians K.J.;
RT   "Mutational analysis of the primosome assembly sites: I. Distinct
RT   classes of mutants in the pBR322 Escherichia coli factor Y DNA
RT   effector sequences";
RL   J. Biol. Chem. 259:14286-14292(1984).
XX
RN   [19]
RP   2348-2415
RX   MEDLINE; 85054886.
RA   Soeller W., Abarzua P.I., Marians K.J.;
RT   "Mutational analysis of primosome assembly sites: II. Role of
RT   secondary structure in the formation of active sites";
RL   J. Biol. Chem. 259:14293-14300(1984).
XX
RN   [20]
RP   1-4361
RX   MEDLINE; 84300294.
RA   Van Dyke M.M., Dervan P.B.;
RT   "Echinomycin binding sites on DNA";
RL   Science 225:1122-1127(1984).
XX
RN   [21]
RA   Pouwels P.H., Enger-Valk B.E., Brammar W.J.;
RT   "Vector I-A-iv-1";
RL   (in) Brammar W.J. (eds.);
RL   CLONING VECTORS:1-1;
RL   Elsevier Scientific Publishing, Amsterdam (1985)
XX
RN   [22]
RP   1-4361
RX   MEDLINE; 89108024.
RA   Watson N.;
RT   "A new revision of the sequence of plasmid pBR322";
RL   Gene 70:399-403(1988).
XX
RN   [23]
RA   Gilbert W.;
RT   "Obtained from VecBase 3.0";
RL   Unpublished.
XX
CC   The circular sequence is numbered such that 0 is the middle of the
CC   unique EcoRI site and the count increases first through the tet
CC   genes, the pMB1 material, and finally through the Tn3 region.
CC   Plasmid pBR322 contains ampicillin and tetracycline resistance
CC   genes. The ampicillin resistance gene (amp-r) is a penicillin
CC   beta-lactamase. Promoters P1 and P3 are for the beta-lactamase
CC   gene. P3 is the natural promoter, and P1 is artificially created by
CC   the ligation of two different DNA fragments to create pBR322. P2 is
CC   in the same region as P1, but it is on the opposite strand and
CC   initiates transcription in the direction of the tetracycline
CC   resistance gene. Mutational studies in the primosome assembly sites
CC   indicate four types of mutations: Class I having no effect on the
CC   activities elicited by the DNA site and the bases involved are
CC   probably spacers; Class II requiring higher Mg-2+ concentrations
CC   than the wild-type to be fully activated as factor Y ATPase
CC   effectors; Class III co-inactivating both the ATPase effector and
CC   DNA replication template activity of the site, indicating that they
CC   probably represent essential contact points between factor Y and
CC   the DNA; Class IV having a replication template activity
CC   intermediate that of class III and class II mutant DNAs. Specific
CC   sites within or near the origins of replication are recognized by
CC   dnaA protein. Without dnaA binding to the origin of replication
CC   chromosomal replication is not possible [15]. pBR322 DNA contains
CC   two separate regions on opposite strands and close to the origin of
CC   replication which, when in single-stranded form, can act as
CC   effectors for the ATPase activity of E.coli replication factor Y
CC   [5]. Small fragments of DNA containing these sites when cloned in
CC   an f1 phage vector act as origins of DNA replication allowing the
CC   formation of complementary double-stranded DNA in
CC   rifampicin-resistant, dna[B,G,C]-dependent fashion in vitro [5].
CC   The biological activity of echinomycin is thought to be related to
CC   the formation of complexes by intercalating with cellular DNA [20].
CC   Complete source information: Plasmid pBR322 from E.coli
CC   [2],[1],[3],[6],[11],[8],[5],[7],[12],
CC   [13],[10],[9],[14],[18],[19],[15],[20],[16]; pBR322 DNA in pXf3
CC   [4]. The following data and their annotation were supplied by Will
CC   Gilbert under the auspices of the Curator Program. CROSSREFERENCE
CC   #parent GenBank(50):pSC101C, GenBank(50):Trn3 #offspring
CC   VecBase(3):pBR325, VecBase(3):pBR327, VecBase(3): pBR328,
CC   VecBase(3):pAT153, VecBase(3):pUC7, VecBase(3): pJRD158,
CC   VecBase(3):PiVX, VecBase(3):PiAN7, VecBase(3): pSP64,
CC   VecBase(3):pSP65, VecBase(3):pGEM1, VecBase(3): pGEM2,
CC   VecBase(3):pGEM3, VecBase(3):pGEM4, VecBase(3): pKK223,
CC   VecBase(3):pLBU3, VecBase(3):pTrS3, VecBase(3): pRSVNeo,
CC   VecBase(3):pSV2Cat, VecBase(3):M13mp9, VecBase(3): pHC79,
CC   VecBase(3):pV34, VecBase(3):pKTH601, VecBase(3): pKTH604,
CC   VecBase(3):pKTH605, VecBase(3):pKTH606, VecBase(3):YEp24,
CC   VecBase(3):YIp5, VecBase(3):YRp17, VecBase(3): pSP18,
CC   VecBase(3):pSP19, VecBase(3):pSP6T3, VecBase(3):pSP6T719,
CC   VecBase(3):pT712, VecBase(3):pT713, VecBase(3):pT7T318, VecBase(3):
CC   pT7T319, VecBase(3):pT7T3A18, VecBase(3):pT7T3A19, VecBase(3):pEX1,
CC   VecBase(3):pEX2, VecBase(3):pEX3, VecBase(3): pCKSP6,
CC   VecBase(3):pACYC177, VecBase(3):pKO1, VecBase(3): pKO2,
CC   VecBase(3):pKM1, VecBase(3):pKM2, VecBase(3):pMBL1, VecBase(3):
CC   pMBL604, VecBase(3):pMC1511, VecBase(3):pMC1871, VecBase(3):pAA37X,
CC   VecBase(3):pUR278, VecBase(3):pUR288, VecBase(3): pUR289,
CC   VecBase(3):pUR290, VecBase(3):pUR291, VecBase(3): pUR292,
CC   VecBase(3):pUR222. NCBI gi: 208958
XX
FH   Key             Location/Qualifiers
FH
FT   source          1..4361
FT                   /organism="Cloning vector"
FT                   /sub_species="Cloning vector pBR322"
FT                   /sequenced_mol="DNA"
FT                   /tissue_lib="ATCC 31344, ATCC 37017"
FT   misc_feature    1..1643
FT                   /note="from pSC101 (bp 1-1643)"
FT   misc_binding    24..27
FT                   /bound_moiety="echinomycin"
FT   promoter        complement(27..33)
FT                   /note="promoter P1 [6]"
FT   misc_binding    39..42
FT                   /bound_moiety="echinomycin"
FT   promoter        43..49
FT                   /note="promoter P2 [6]"
FT   misc_binding    53..56
FT                   /bound_moiety="echinomycin"
FT   misc_binding    67..70
FT                   /bound_moiety="echinomycin"
FT   misc_binding    80..83
FT                   /bound_moiety="echinomycin"
FT   CDS             86..1276
FT                   /gene="tet"
FT                   /note="NCBI gi: 208959"
FT                   /codon_start=1
FT                   /product="tetracycline resistance protein"
FT                   /db_xref="PID:g208959"
FT                   /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
FT                   GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
FT                   VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
FT                   LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
FT                   QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
FT                   AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
FT                   AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
FT   misc_feature    complement(141..142)
FT                   /note="Endo.Sce I cleavage site coordinated with site at
FT                   base 146 [10]"
FT   misc_feature    146..147
FT                   /note="Endo.Sce I cleavage site coordinated with site at
FT                   base 142 [10]"
FT   misc_binding    411..414
FT                   /bound_moiety="echinomycin"
FT   conflict        replace(426,"")
FT                   /citation=[11]
FT   misc_binding    469..472
FT                   /bound_moiety="echinomycin"
FT   old_sequence    526..528
FT                   /citation=[17]
FT   repeat_unit     complement(1515..1519)
FT                   /note="gamma-delta insertion target sequence"
FT                   /rpt_type=DIRECT
FT   misc_feature    1636..1762
FT                   /note="from pSC101 (bp 1860-1986)"
FT   misc_feature    1763..3147
FT                   /note="from pMB1"
FT   repeat_unit     complement(1788..1792)
FT                   /note="gamma-delta insertion target sequence"
FT                   /rpt_type=DIRECT
FT   conflict        replace(1891..1892,"att")
FT                   /citation=[23]
FT   old_sequence    1892..1893
FT                   /citation=[2]
FT                   /citation=[22]
FT   RBS             1905..1910
FT   RBS             1905..1909
FT                   /note="Shine-Dalgarno sequence"
FT   conflict        replace(1913..1914,"caa")
FT                   /citation=[23]
FT   old_sequence    1914..1915
FT                   /citation=[17]
FT   CDS             1915..2106
FT                   /note="NCBI gi: 456436"
FT                   /codon_start=1
FT                   /product="ROP protein"
FT                   /db_xref="PID:g456436"
FT                   /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
FT                   DELYRSCLARFGDDGENL"
FT   CDS             1915..2106
FT                   /note="NCBI gi: 506846"
FT                   /codon_start=1
FT                   /product="ROP protein"
FT                   /db_xref="PID:g506846"
FT                   /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
FT                   DELYRSCLARFGDDGENL"
FT   misc_feature    2011..2167
FT                   /note="H-strand Y effector site [5]"
FT   repeat_unit     complement(2245..2249)
FT                   /note="gamma-delta insertion target sequence"
FT                   /rpt_type=DIRECT
FT   misc_feature    complement(2351..2414)
FT                   /note="L-strand Y effector site [5]"
FT   misc_binding    2439..2447
FT                   /bound_moiety="dnaA"
FT   rep_origin      2535
FT   old_sequence    replace(2729..2730,"at")
FT                   /note="revision according to [17]"
FT                   /citation=[17]
FT                   /citation=[2]
FT   old_sequence    2729
FT                   /citation=[17]
FT   old_sequence    replace(2730,"t")
FT                   /note="revision according to [16]"
FT                   /citation=[2]
FT                   /citation=[16]
FT   old_sequence    2730
FT                   /citation=[17]
FT   misc_feature    3148..4361
FT                   /note="from Tn3 (bp 4957-3742)"
FT   repeat_region   3148..3185
FT                   /note="corresponds to one of the 38bp repeats found in Tn3
FT                   (bp 1-38 and complement (4920-4957))"
FT                   /rpt_type=INVERTED
FT   CDS             complement(3293..4153)
FT                   /gene="bla"
FT                   /note="E-286; NCBI gi: 455370"
FT                   /codon_start=1
FT                   /product="beta-lactamase"
FT                   /db_xref="PID:g455370"
FT                   /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
FT                   ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
FT                   PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
FT                   EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
FT                   LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
FT                   LIKHW"
FT   mat_peptide     complement(3296..4084)
FT                   /gene="bla"
FT                   /codon_start=1
FT                   /product="beta-lactamase"
FT   sig_peptide     complement(4085..4153)
FT                   /gene="bla"
FT                   /codon_start=1
FT   RBS             complement(4161..4165)
FT                   /note="Shine-Dalgarno sequence"
FT   promoter        complement(4188..4194)
FT                   /note="promoter P3 [6]"
FT   misc_binding    complement(4268..4271)
FT                   /bound_moiety="echinomycin"
FT   misc_binding    complement(4280..4283)
FT                   /bound_moiety="echinomycin"
FT   misc_binding    complement(4285..4288)
FT                   /bound_moiety="echinomycin"
FT   misc_binding    complement(4296..4299)
FT                   /bound_moiety="echinomycin"
FT   misc_binding    complement(4311..4314)
FT                   /bound_moiety="echinomycin"
FT   misc_binding    complement(4317..4320)
FT                   /bound_moiety="echinomycin"
FT   misc_binding    complement(4331..4334)
FT                   /bound_moiety="echinomycin"
XX
SQ   Sequence 4361 BP; 983 A; 1210 C; 1134 G; 1034 T; 0 other;
     ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa        60
     ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg       120
     caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt       180
     gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata       240
     tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg       300
     ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac       360
     cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac       420
     aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca       480
     cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg       540
     actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct       600
     caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat       660
     gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt       720
     cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct       780
     ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct       840
     tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa       900
     acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt       960
     cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc      1020
     cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca      1080
     tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc      1140
     gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat      1200
     tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg      1260
     ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga      1320
     attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac      1380
     atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg      1440
     tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg      1500
     ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc      1560
     tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg      1620
     taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca      1680
     ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga      1740
     ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa      1800
     cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt      1860
     ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc      1920
     aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt      1980
     ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac      2040
     gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac      2100
     ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc      2160
     agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc      2220
     cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg      2280
     tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc      2340
     gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc      2400
     ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata      2460
     acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg      2520
     cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct      2580
     caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa      2640
     gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc      2700
     tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt      2760
     aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg      2820
     ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg      2880
     cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct      2940
     tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc      3000
     tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg      3060
     ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc      3120
     aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt      3180
     aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa      3240
     aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat      3300
     gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct      3360
     gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg      3420
     caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag      3480
     ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta      3540
     attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg      3600
     ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg      3660
     gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct      3720
     ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta      3780
     tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg      3840
     gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc      3900
     cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg      3960
     gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga      4020
     tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg      4080
     ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat      4140
     gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc      4200
     tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca      4260
     catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct      4320
     ataaaaatag gcgtatcacg aggccctttc gtcttcaaga a                          4361
//