LOCUS       Exported               10867 bp ds-DNA     circular SYN 05-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10867)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pENTR taken from kinase library, KIN_482

            

            in-silico sequence added (B1 - insert your ORF - B2)

            #######################

            

            Open pcDNA3_X-Glink-TEV1-118-tevS-GV-2xHA with BmgBI,

            insert RFB. (our RfB: TAT --> cta (pos 125-127)

            

            

            Digest with EcoRI for orientation: 6330 + 2163 bp

            

            #############

            insert BamHI - BmgBI - GGGGS-2xlinker - NTEV - tevS- GV - 2xHA - 
            Stop - ApaI int

            

            PCR on NTEV-tevS-GV:

            SE: 
            GAGAGGATCCCACGTCGGAGGTGGAGGTAGCGGAGGTGGAGGTAGCGGAGAGAGCTTGTTTAAGGGAC
            CAC

            AS: 
            TCTCGGGCCCTCATGCATAGTCCGGGACGTCATAGGGATAGCCCGCATAGTCAGGAACATCGTATGGG
            TAGCCCCCACCGTACTCGTCAATTCCAAG

            

            CTTGGAATTGACGAGTACGGTGGGGGCTACCCATACGATGTTCCTGACTATGCGGGCTATCCCTATGA
            CGTCCCGGACTATGCATGAGGGCCCGAGA

            

            PCR as adapter: 243-mNTEV-GV_AS_adapter: 
            GGATAGCCCGCATAGTCAGGAACATCGTATGGGTAGCCCCCACCGTACTCGTCAATTCCAAG

            

            

            ####################################################################
            #

            History of cloning:

            

            GVmut inserted with XbaI + ApaI

            

            ####################################################################
            #

            

            Gal4-VP16 ORF: Site directed mutagenesis of BmgBI (site on reverse 
            strand!!!). GACGTG --> GACGTc (pos. 2088)

            SE: GCGAGGACGTCGCGATGGCG

            AS: CGCCATCGCGACGTCCTCGC

            

            

            #############################################################

            PCR for XbaI - GV - Stop - ApaI

            

            PCR on GV:

            SE: GAGATCTAGAGAACAGAAGCTGATCTCAGAAGAGGATC

            AS: TCTCGGGCCCTTAATAATAAAAATCATAAATCATAAGACATTCGCCCGG

            

            AS: 
            TCTCGGGCCCCTACTTATCGTCGTCATCCTTGTAATCATAATAAAAATCATAAATCATAAGACATTCG
            CCCGG  (with Flag-Tag)

            

            CCGGGCGAATGTCTTATGATTTATGATTTTTATTATGATTACAAGGATGACGACGATAAGTAGGGGCC
            CGAGA

            

            

            #############################################################

            PCR for BamHI - BmgBI - ASPlinker-TEV1-118 - tevS - XbaI

            

            PCR on pcDNA3_X-ASPlink-TEV1-118-Flag_AmpR.

            

            SE: GAGAGGATCCCACGTCGCCAGCCCTAGCAACCCTGG

            AS: 
            TCTCTCTAGAACCCTGGAAGTACAGGTTCTCGGGAGCTTGGAAGTTGGTTGTCACAAGACATATG
FEATURES             Location/Qualifiers
     source          1..10867
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        209..863
                     /label=CMV promotor
                     /note="CMV promotor"
     primer_bind     863..882
                     /label=T7 promoter/priming site
                     /note="T7 promoter/priming site"
     misc_recomb     920..938
                     /label=attB1
                     /note="attB1"
     CDS             947..6406
                     /codon_start=1
                     /label=ERBB3-NTEV-tcs-GV-2HA
                     /translation="MRANDALQVLGLLFSLARGSEVGNSQAVCPGTLNGLSVTGDAENQ
                     YQTLYKLYERCEVVMGNLEIVLTGHNADLSFLQWIREVTGYVLVAMNEFSTLPLPNLRV
                     VRGTQVYDGKFAIFVMLNYNTNSSHALRQLRLTQLTEILSGGVYIEKNDKLCHMDTIDW
                     RDIVRDRDAEIVVKDNGRSCPPCHEVCKGRCWGPGSEDCQTLTKTICAPQCNGHCFGPN
                     PNQCCHDECAGGCSGPQDTDCFACRHFNDSGACVPRCPQPLVYNKLTFQLEPNPHTKYQ
                     YGGVCVASCPHNFVVDQTSCVRACPPDKMEVDKNGLKMCEPCGGLCPKACEGTGSGSRF
                     QTVDSSNIDGFVNCTKILGNLDFLITGLNGDPWHKIPALDPEKLNVFRTVREITGYLNI
                     QSWPPHMHNFSVFSNLTTIGGRSLYNRGFSLLIMKNLNVTSLGFRSLKEISAGRIYISA
                     NRQLCYHHSLNWTKVLRGPTEERLDIKHNRPRRDCVAEGKVCDPLCSSGGCWGPGPGQC
                     LSCRNYSRGGVCVTHCNFLNGEPREFAHEAECFSCHPECQPMEGTATCNGSGSDTCAQC
                     AHFRDGPHCVSSCPHGVLGAKGPIYKYPDVQNECRPCHENCTQGCKGPELQDCLGQTLV
                     LIGKTHLTMALTVIAGLVVIFMMLGGTFLYWRGRRIQNKRAMRRYLERGESIEPLDPSE
                     KANKVLARIFKETELRKLKVLGSGVFGTVHKGVWIPEGESIKIPVCIKVIEDKSGRQSF
                     QAVTDHMLAIGSLDHAHIVRLLGLCPGSSLQLVTQYLPLGSLLDHVRQHRGALGPQLLL
                     NWGVQIAKGMYYLEEHGMVHRNLAARNVLLKSPSQVQVADFGVADLLPPDDKQLLYSEA
                     KTPIKWMALESIHFGKYTHQSDVWSYGVTVWELMTFGAEPYAGLRLAEVPDLLEKGERL
                     AQPQICTIDVYMVMVKCWMIDENIRPTFKELANEFTRMARDPPRYLVIKRESGPGIAPG
                     PEPHGLTNKKLEEVELEPELDLDLDLEAEEDNLATTTLGSALSLPVGTLNRPRGSQSLL
                     SPSSGYMPMNQGNLGESCQESAVSGSSERCPRPVSLHPMPRGCLASESSEGHVTGSEAE
                     LQEKVSMCRSRSRSRSPRPRGDSAYHSQRHSLLTPVTPLSPPGLEEEDVNGYVMPDTHL
                     KGTPSSREGTLSSVGLSSVLGTEEEDEDEEYEYMNRRRRHSPPHPPRPSSLEELGYEYM
                     DVGSDLSASLGSTQSCPLHPVPIMPTAGTTPDEDYEYMNRQRDGGGPGGDYAAMGACPA
                     SEQGYEEMRAFQGPGHQAPHVHYARLKTLRSLEATDSAFDNPDYWHSRLFPKANAQRTD
                     PAFLYKVVDVGGGGSGGGGSGESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGP
                     FIITNKHLFRRNNGTLLVQSLHGVFKVKDTTTLQQHLIDGRDMMIIRMPKDFPPFPQKL
                     KFREPQREERICLVTTNFQAPENLYFQGSREQKLISEEDLARAMKLLSSIEQACDICRL
                     KKLKCSKEKPKCAKCLKNNWECRYSPKTKRSPLTRAHLTEVESRLERLEQLFLLIFPRE
                     DLDMILKMDSLQDIKALLTGLFVQDNVNKDAVTDRLASVETDMPLTLRQHRISATSSSE
                     ESSNKGQRQLTVSPEFPGISLIIRFTMPKKKRKVAYSRARTKNNYGSTIEGLLDLPDDD
                     APEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHADALDDFDLDM
                     LGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGGGYPYDVPDYAGYPYD
                     VPDYA"
     CDS             947..4972
                     /codon_start=1
                     /label=receptor tyrosine-protein kinase erbB-3
                     /note="isoform 1 precursor is encoded by transcript variant
                     1; receptor tyrosine-protein kinase erbB-3; 
                     proto-oncogene-like protein c-ErbB-3; tyrosine kinase-type 
                     cell surface receptor HER3"
                     /translation="MRANDALQVLGLLFSLARGSEVGNSQAVCPGTLNGLSVTGDAENQ
                     YQTLYKLYERCEVVMGNLEIVLTGHNADLSFLQWIREVTGYVLVAMNEFSTLPLPNLRV
                     VRGTQVYDGKFAIFVMLNYNTNSSHALRQLRLTQLTEILSGGVYIEKNDKLCHMDTIDW
                     RDIVRDRDAEIVVKDNGRSCPPCHEVCKGRCWGPGSEDCQTLTKTICAPQCNGHCFGPN
                     PNQCCHDECAGGCSGPQDTDCFACRHFNDSGACVPRCPQPLVYNKLTFQLEPNPHTKYQ
                     YGGVCVASCPHNFVVDQTSCVRACPPDKMEVDKNGLKMCEPCGGLCPKACEGTGSGSRF
                     QTVDSSNIDGFVNCTKILGNLDFLITGLNGDPWHKIPALDPEKLNVFRTVREITGYLNI
                     QSWPPHMHNFSVFSNLTTIGGRSLYNRGFSLLIMKNLNVTSLGFRSLKEISAGRIYISA
                     NRQLCYHHSLNWTKVLRGPTEERLDIKHNRPRRDCVAEGKVCDPLCSSGGCWGPGPGQC
                     LSCRNYSRGGVCVTHCNFLNGEPREFAHEAECFSCHPECQPMEGTATCNGSGSDTCAQC
                     AHFRDGPHCVSSCPHGVLGAKGPIYKYPDVQNECRPCHENCTQGCKGPELQDCLGQTLV
                     LIGKTHLTMALTVIAGLVVIFMMLGGTFLYWRGRRIQNKRAMRRYLERGESIEPLDPSE
                     KANKVLARIFKETELRKLKVLGSGVFGTVHKGVWIPEGESIKIPVCIKVIEDKSGRQSF
                     QAVTDHMLAIGSLDHAHIVRLLGLCPGSSLQLVTQYLPLGSLLDHVRQHRGALGPQLLL
                     NWGVQIAKGMYYLEEHGMVHRNLAARNVLLKSPSQVQVADFGVADLLPPDDKQLLYSEA
                     KTPIKWMALESIHFGKYTHQSDVWSYGVTVWELMTFGAEPYAGLRLAEVPDLLEKGERL
                     AQPQICTIDVYMVMVKCWMIDENIRPTFKELANEFTRMARDPPRYLVIKRESGPGIAPG
                     PEPHGLTNKKLEEVELEPELDLDLDLEAEEDNLATTTLGSALSLPVGTLNRPRGSQSLL
                     SPSSGYMPMNQGNLGESCQESAVSGSSERCPRPVSLHPMPRGCLASESSEGHVTGSEAE
                     LQEKVSMCRSRSRSRSPRPRGDSAYHSQRHSLLTPVTPLSPPGLEEEDVNGYVMPDTHL
                     KGTPSSREGTLSSVGLSSVLGTEEEDEDEEYEYMNRRRRHSPPHPPRPSSLEELGYEYM
                     DVGSDLSASLGSTQSCPLHPVPIMPTAGTTPDEDYEYMNRQRDGGGPGGDYAAMGACPA
                     SEQGYEEMRAFQGPGHQAPHVHYARLKTLRSLEATDSAFDNPDYWHSRLFPKANAQRT"
     sig_peptide     947..1003
                     /label=ERBB3
                     /note="Potential; propagated from UniProtKB/Swiss-Prot 
                     (P21860.1)"
     mat_peptide     1004..4972
                     /label=Receptor tyrosine-protein kinase erbB-3
                     /note="propagated from UniProtKB/Swiss-Prot (P21860.1)"
     exon            1029..1180
                     /label=ERBB3
     exon            1181..1367
                     /label=ERBB3
     exon            1368..1493
                     /label=ERBB3
     exon            1494..1559
                     /label=ERBB3
     exon            1560..1678
                     /label=ERBB3
     exon            1679..1820
                     /label=ERBB3
     exon            1821..1934
                     /label=ERBB3
     exon            1935..2055
                     /label=ERBB3
     exon            2056..2129
                     /label=ERBB3
     exon            2130..2220
                     /label=ERBB3
     exon            2221..2426
                     /label=ERBB3
     exon            2427..2559
                     /label=ERBB3
     exon            2560..2650
                     /label=ERBB3
     exon            2651..2805
                     /label=ERBB3
     exon            2806..2859
                     /label=ERBB3
     exon            2860..3001
                     /label=ERBB3
     misc_feature    2876..2938
                     /label=ERBB3
                     /note="propagated from UniProtKB/Swiss-Prot (P21860.1); 
                     transmembrane region"
     misc_feature    2984..2986
                     /label=ERBB3
                     /note="Phosphotyrosine; propagated from 
                     UniProtKB/Swiss-Prot (P21860.1); phosphorylation site"
     exon            3002..3121
                     /label=ERBB3
     misc_feature    3002..3004
                     /label=ERBB3
                     /note="Phosphoserine; propagated from UniProtKB/Swiss-Prot 
                     (P21860.1); phosphorylation site"
     CDS             3071..3844
                     /codon_start=1
                     /label=kinase domain
                     /note="kinase domain"
                     /translation="LRKLKVLGSGVFGTVHKGVWIPEGESIKIPVCIKVIEDKSGRQSF
                     QAVTDHMLAIGSLDHAHIVRLLGLCPGSSLQLVTQYLPLGSLLDHVRQHRGALGPQLLL
                     NWGVQIAKGMYYLEEHGMVHRNLAARNVLLKSPSQVQVADFGVADLLPPDDKQLLYSEA
                     KTPIKWMALESIHFGKYTHQSDVWSYGVTVWELMTFGAEPYAGLRLAEVPDLLEKGERL
                     AQPQICTIDVYMVMVKCWMIDENIRPTFKELANEFT"
     exon            3122..3220
                     /label=ERBB3
     exon            3221..3406
                     /label=ERBB3
     exon            3407..3562
                     /label=ERBB3
     exon            3563..3638
                     /label=ERBB3
     exon            3639..3785
                     /label=ERBB3
     exon            3786..3883
                     /label=ERBB3
     exon            3884..4075
                     /label=ERBB3
     exon            4076..4147
                     /label=ERBB3
     exon            4148..4448
                     /label=ERBB3
     misc_feature    4928..4930
                     /label=ERBB3
                     /note="Phosphotyrosine; propagated from 
                     UniProtKB/Swiss-Prot (P21860.1); phosphorylation site"
     misc_recomb     4974..4998
                     /label=attB2
                     /note="attB2"
     CDS             5006..5035
                     /codon_start=1
                     /label=GS linker
                     /note="GS linker"
                     /translation="GGGGSGGGGS"
     CDS             5036..5389
                     /codon_start=1
                     /label=TEV1-118
                     /note="TEV1-118"
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKDTTTLQQHLIDGRDMMIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQA"
     misc_binding    complement(5269..5291)
                     /label=NTEV 5' sequencing primer
                     /note="NTEV 5' sequencing primer"
     CDS             5393..5413
                     /codon_start=1
                     /label=tevS
                     /note="tevS (TEV cleavage site)"
                     /translation="ENLYFQG"
     CDS             5420..5449
                     /codon_start=1
                     /label=Myc-tag
                     /note="Myc-tag (due to cloning reasons - taken from old 
                     pDEST_X-ASPlink-NTEV-tevS-GV which contained that Myc-tag 
                     already)"
                     /translation="EQKLISEEDL"
     CDS             5450..6346
                     /codon_start=1
                     /label=Gal4-VP16
                     /note="Gal4-VP16"
                     /translation="ARAMKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSP
                     KTKRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDN
                     VNKDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSPEFPGISLIIRFT
                     MPKKKRKVAYSRARTKNNYGSTIEGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTA
                     PPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMAD
                     FEFEQMFTDALGIDEYGG"
     CDS             6347..6406
                     /codon_start=1
                     /label=2xHA-tag
                     /note="2xHA-tag"
                     /translation="GYPYDVPDYAGYPYDVPDYA"
     polyA_site      6439..6670
                     /label=BGH polyA
                     /note="BGH polyA"
     primer_bind     6457..6474
                     /label=BGH Reverse priming site
                     /note="BGH Reverse priming site"
     rep_origin      6750..7056
                     /label=f1 origin
                     /note="f1 origin"
     promoter        7211..7536
                     /label=SV40 promotor
                     /note="SV40 promotor"
     CDS             7572..8366
                     /codon_start=1
                     /label=Neomycin resistance gene
                     /note="Neomycin resistance gene"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    8421..8793
                     /label=SV40 poly A
                     /note="SV40 poly A"
     rep_origin      9053..9726
                     /direction=RIGHT
                     /label=ColE1 origin (pBR3222 origin)
                     /note="ColE1 origin (pBR3222 origin)"
     CDS             complement(9871..10731)
                     /codon_start=1
                     /label=AmpR
                     /note="AmpR"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cccacatcaa caagtttgta caaaaaagtt ggcaccatga gggcgaacga
      961 cgctctgcag gtgctgggct tgcttttcag cctggcccgg ggctccgagg tgggcaactc
     1021 tcaggcagtg tgtcctggga ctctgaatgg cctgagtgtg accggcgatg ctgagaacca
     1081 ataccagaca ctgtacaagc tctacgagag gtgtgaggtg gtgatgggga accttgagat
     1141 tgtgctcacg ggacacaatg ccgacctctc cttcctgcag tggattcgag aagtgacagg
     1201 ctatgtcctc gtggccatga atgaattctc tactctacca ttgcccaacc tccgcgtggt
     1261 gcgagggacc caggtctacg atgggaagtt tgccatcttc gtcatgttga actataacac
     1321 caactccagc cacgctctgc gccagctccg cttgactcag ctcaccgaga ttctgtcagg
     1381 gggtgtttat attgagaaga acgataagct ttgtcacatg gacacaattg actggaggga
     1441 catcgtgagg gaccgagatg ctgagatagt ggtgaaggac aatggcagaa gctgtccccc
     1501 ctgtcatgag gtttgcaagg ggcgatgctg gggtcctgga tcagaagact gccagacatt
     1561 gaccaagacc atctgtgctc ctcagtgtaa tggtcactgc tttgggccca accccaacca
     1621 gtgctgccat gatgagtgtg ccgggggctg ctcaggccct caggacacag actgctttgc
     1681 ctgccggcac ttcaatgaca gtggagcctg tgtacctcgc tgtccacagc ctcttgtcta
     1741 caacaagcta actttccagc tggaacccaa tccccacacc aagtatcagt atggaggagt
     1801 ttgtgtagcc agctgtcccc ataactttgt ggtggatcaa acatcctgtg tcagggcctg
     1861 tcctcctgac aagatggaag tagataaaaa tgggctcaag atgtgtgagc cttgtggggg
     1921 actatgtccc aaagcctgtg agggaacagg ctctgggagc cgcttccaga ctgtggactc
     1981 gagcaacatt gatggatttg tgaactgcac caagatcctg ggcaacctgg actttctgat
     2041 caccggcctc aatggagacc cctggcacaa gatccctgcc ctggacccag agaagctcaa
     2101 tgtcttccgg acagtacggg agatcacagg ttacctgaac atccagtcct ggccgcccca
     2161 catgcacaac ttcagtgttt tttccaattt gacaaccatt ggaggcagaa gcctctacaa
     2221 ccggggcttc tcattgttga tcatgaagaa cttgaatgtc acatctctgg gcttccgatc
     2281 cctgaaggaa attagtgctg ggcgtatcta tataagtgcc aataggcagc tctgctacca
     2341 ccactctttg aactggacca aggtgcttcg ggggcctacg gaagagcgac tagacatcaa
     2401 gcataatcgg ccgcgcagag actgcgtggc agagggcaaa gtgtgtgacc cactgtgctc
     2461 ctctggggga tgctggggcc caggccctgg tcagtgcttg tcctgtcgaa attatagccg
     2521 aggaggtgtc tgtgtgaccc actgcaactt tctgaatggg gagcctcgag aatttgccca
     2581 tgaggccgaa tgcttctcct gccacccgga atgccaaccc atggagggca ctgccacatg
     2641 caatggctcg ggctctgata cttgtgctca atgtgcccat tttcgagatg ggccccactg
     2701 tgtgagcagc tgcccccatg gagtcctagg tgccaagggc ccaatctaca agtacccaga
     2761 tgttcagaat gaatgtcggc cctgccatga gaactgcacc caggggtgta aaggaccaga
     2821 gcttcaagac tgtttaggac aaacactggt gctgatcggc aaaacccatc tgacaatggc
     2881 tttgacagtg atagcaggat tggtagtgat tttcatgatg ctgggcggca cttttctcta
     2941 ctggcgtggg cgccggattc agaataaaag ggctatgagg cgatacttgg aacggggtga
     3001 gagcatagag cctctggacc ccagtgagaa ggctaacaaa gtcttggcca gaatcttcaa
     3061 agagacagag ctaaggaagc ttaaagtgct tggctcgggt gtctttggaa ctgtgcacaa
     3121 aggagtgtgg atccctgagg gtgaatcaat caagattcca gtctgcatta aagtcattga
     3181 ggacaagagt ggacggcaga gttttcaagc tgtgacagat catatgctgg ccattggcag
     3241 cctggaccat gcccacattg taaggctgct gggactatgc ccagggtcat ctctgcagct
     3301 tgtcactcaa tatttgcctc tgggttctct gctggatcat gtgagacaac accggggggc
     3361 actggggcca cagctgctgc tcaactgggg agtacaaatt gccaagggaa tgtactacct
     3421 tgaggaacat ggtatggtgc atagaaacct ggctgcccga aacgtgctac tcaagtcacc
     3481 cagtcaggtt caggtggcag attttggtgt ggctgacctg ctgcctcctg atgataagca
     3541 gctgctatac agtgaggcca agactccaat taagtggatg gcccttgaga gtatccactt
     3601 tgggaaatac acacaccaga gtgatgtctg gagctatggt gtgacagttt gggagttgat
     3661 gaccttcggg gcagagccct atgcagggct acgattggct gaagtaccag acctgctaga
     3721 gaagggggag cggttggcac agccccagat ctgcacaatt gatgtctaca tggtgatggt
     3781 caagtgttgg atgattgatg agaacattcg cccaaccttt aaagaactag ccaatgagtt
     3841 caccaggatg gcccgagacc caccacggta tctggtcata aagagagaga gtgggcctgg
     3901 aatagcccct gggccagagc cccatggtct gacaaacaag aagctagagg aagtagagct
     3961 ggagccagaa ctagacctag acctagactt ggaagcagag gaggacaacc tggcaaccac
     4021 cacactgggc tccgccctca gcctaccagt tggaacactt aatcggccac gtgggagcca
     4081 gagcctttta agtccatcat ctggatacat gcccatgaac cagggtaatc ttggggagtc
     4141 ttgccaggag tctgcagttt ctgggagcag tgaacggtgc ccccgtccag tctctctaca
     4201 cccaatgcca cggggatgcc tggcatcaga gtcatcagag gggcatgtaa caggctctga
     4261 ggctgagctc caggagaaag tgtcaatgtg taggagccgg agcaggagcc ggagcccacg
     4321 gccacgcgga gatagcgcct accattccca gcgccacagt ctgctgactc ctgttacccc
     4381 actctcccca cccgggttag aggaagagga tgtcaacggt tatgtcatgc cagatacaca
     4441 cctcaaaggt actccctcct cccgggaagg caccctttct tcagtgggtc tcagttctgt
     4501 cctgggtact gaagaagaag atgaagatga ggagtatgaa tacatgaacc ggaggagaag
     4561 gcacagtcca cctcatcccc ctaggccaag ttcccttgag gagctgggtt atgagtacat
     4621 ggatgtgggg tcagacctca gtgcctctct gggcagcaca cagagttgcc cactccaccc
     4681 tgtacccatc atgcccactg caggcacaac tccagatgaa gactatgaat atatgaatcg
     4741 gcaacgagat ggaggtggtc ctgggggtga ttatgcagcc atgggggcct gcccagcatc
     4801 tgagcaaggg tatgaagaga tgagagcttt tcaggggcct ggacatcagg ccccccatgt
     4861 ccattatgcc cgcctaaaaa ctctacgtag cttagaggct acagactctg cctttgataa
     4921 ccctgattac tggcatagca ggcttttccc caaggctaat gcccagagaa cggacccagc
     4981 tttcttgtac aaagtggttg atgtcggagg tggaggtagc ggaggtggag gtagcggaga
     5041 gagcttgttt aagggaccac gtgattacaa cccgatatcg agcaccattt gtcatttgac
     5101 gaatgaatct gatgggcaca caacatcgtt gtatggtatt ggatttggtc ccttcatcat
     5161 tacaaacaag cacttgtttc gtcgcaataa tggaacactg ttggtccaat cactacatgg
     5221 tgtattcaag gtcaaggaca ccacgacttt gcaacaacac ctcattgatg ggagggacat
     5281 gatgattatt cgcatgccta aggatttccc accatttcct caaaagctga aatttagaga
     5341 gccacaaagg gaagagcgca tatgtcttgt gacaaccaac ttccaagctc ccgagaacct
     5401 gtacttccag ggttctagag aacagaagct gatctcagaa gaggatctgg cccgggccat
     5461 gaagctactg tcttctatcg aacaagcatg cgatatttgc cgacttaaaa agctcaagtg
     5521 ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac aactgggagt gtcgctactc
     5581 tcccaaaacc aaaaggtctc cgctgactag ggcacatctg acagaagtgg aatcaaggct
     5641 agaaagactg gaacagctat ttctactgat ttttcctcga gaagaccttg acatgatttt
     5701 gaaaatggat tctttacagg atataaaagc attgttaaca ggattatttg tacaagataa
     5761 tgtgaataaa gatgccgtca cagatagatt ggcttcagtg gagactgata tgcctctaac
     5821 attgagacag catagaataa gtgcgacatc atcatcggaa gagagtagta acaaaggtca
     5881 aagacagttg actgtatcgc cggaattccc ggggatcagc ttgataattc gattcaccat
     5941 gcccaagaag aagaggaagg tggcgtacag ccgcgcgcgt acgaaaaaca attacgggtc
     6001 taccatcgag ggcctgctcg atctcccgga cgacgacgcc cccgaagagg cggggctggc
     6061 ggctccgcgc ctgtcctttc tccccgcggg acacacgcgc agactgtcga cggccccccc
     6121 gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtcg cgatggcgca
     6181 tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc
     6241 gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg acttcgagtt
     6301 tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtgggggct acccatacga
     6361 tgttcctgac tatgcgggct atccctatga cgtcccggac tatgcatgag ggccctattc
     6421 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     6481 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     6541 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     6601 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     6661 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     6721 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6781 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6841 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag
     6901 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6961 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7021 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     7081 cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt
     7141 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     7201 cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     7261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     7321 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     7381 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     7441 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     7501 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     7561 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     7621 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     7681 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     7741 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     7801 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     7861 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     7921 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     7981 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     8041 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     8101 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     8161 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     8221 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     8281 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     8341 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     8401 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     8461 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     8521 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     8581 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     8641 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     8701 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     8761 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     8821 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     8881 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     8941 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     9001 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     9061 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     9121 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     9181 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     9241 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     9301 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     9361 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     9421 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     9481 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     9541 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     9601 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     9661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     9781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     9841 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     9901 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     9961 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    10021 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    10081 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    10141 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    10201 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    10261 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    10321 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    10381 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    10441 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    10501 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    10561 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    10621 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    10681 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    10741 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    10801 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    10861 tgacgtc
//
