LOCUS       sequence-343488-       16594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  doxycycline inducible BE3 expression for human CLYBL locus knock-in.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16594)
  AUTHORS   Coelho MA, Cooper S, Strauss ME, Karakoc E, Bhosle S, Goncalves E, 
            Picco G, Burgold T, Cattaneo CM, Veninga V, Consonni S, Dincer C, 
            Vieira SF, Gibson F, Barthorpe S, Hardy C, Rein J, Thomas M, Marioni
            J, Voest EE, Bassett A, Garnett MJ
  TITLE     Base editing screens map mutations affecting interferon-gamma 
            signaling in cancer.
  JOURNAL   Cancer Cell. 2023 Feb 13;41(2):288-303.e6. doi: 
            10.1016/j.ccell.2022.12.009. Epub 2023 Jan 19.
  PUBMED    36669486
REFERENCE   2  (bases 1 to 16594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2022.12.009"; journalName: "Cancer Cell"; date: 
            "2023-02-13- 13"; volume: "41"; issue: "2"; pages: "288-303.e6"
COMMENT     Sequence Label: iBE3 CLYBL
FEATURES             Location/Qualifiers
     source          1..16594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     266..285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2052..2429
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2059..2077
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2095..2113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2131..2149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2167..2185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2203..2221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2239..2257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2275..2293
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2338..2362
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2450..2470
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2471..3154
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             join(3203..3226,3227..3229,3230..7303)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             7216..7224
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7316..7564
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7577..7597
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7604..7624
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7657..7778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7694..7713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7748..7767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7827..7843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7827..7843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7840..7862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7851..7867
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7875..7881,7882..7899,7900..7905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7920..7941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8058..8075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8367..8386
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    8385..8440
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     8486..8505
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(8596..9342)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(9374..9393)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          9401..10417
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(9455..9477)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        10697..11076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    11082..11306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     11295..11312
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    11328..11361
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      12139..12148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(12197..12223)
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             complement(12239..12265)
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             complement(12272..12325)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     LTR             12794..13062
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(13075..13286)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    13322..13546
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(13551..14351)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(13662..13681)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     14272..14291
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(14346..14399)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    14423..14448
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    14449..14482
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(15340..15408,15409..16200)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(15558..15577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(16371..16594,1..365)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
       61 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc gaattcgccc ttagagcaga
      421 gccaggaacc cctgtaggga aggggcagga gagccagggg catgagatgg tggacgagga
      481 agggggacag ggaagcctga gcgcctctcc tgggcttgcc aaggactcaa acccagaagc
      541 ccagagcagg gccttaggga agcgggacgg ctctgttgga gagtccagta ttgaattagt
      601 gatagataca cagaaaaaaa gcaaattgaa taggtgaatg gataaacaaa ctgtggaaca
      661 tccagacaat gaaatattca ccaataaaaa gaaatgagct atcaagccac attggctatg
      721 cccttcacat ttaaggagga accttaaatg catattgcca aattttaaaa gctagtctga
      781 aaatgctaca gactgtatga ttccaattat atgacatttt agaaaaggca aaactataga
      841 gatcagtggt tgccagcggc tcatggaggc aggatgaatt ggtggaacac agggatgctt
      901 tcagggtaat gaaaatatct tgtgtgatcc tgtaataatg gatacgtggc ataatgcatt
      961 tgttaaaacc tgtaaactag ccaggtatag tggctttttt gcacctatag tctcagctac
     1021 ttggtaggct gagacaggaa gatcacttga gcccaggact tcaagaccag cctgggtaat
     1081 atagtaagac cccatctcta ttttttaaag aaattaagaa agaaaagaaa cccatcaact
     1141 gtataacaca aagaatcgta atgtaaacta cagtcctgag taatgaaaca tttttaaaga
     1201 aataaataaa ttcaaggcaa ttattaactt caggaaaaac agtaaagtaa acataagtat
     1261 acaaatcaaa catggctcag ttgtgaaaat atttacaagg tcactttaag taacattgaa
     1321 tgttgatata atcccaaaat atatttatgt tggaaggatg aggagattta attaagggag
     1381 gtggtccctg cagttacgcc aatgataacc cccgccagaa aaatcttagt agccttccct
     1441 ttttgttttc cgtgccccaa ctcggcggat tgactcggcc ccttccggaa acacccgaat
     1501 caacttctag tcaaattatt gttcacgccg caatgaccca cccctggccc gcgtctgtgg
     1561 aactgacccc tggtgtacag gagagttcgc tgctgaaagt ggtcccaaag gggtactagt
     1621 ttttaagctc ccaactcccc ctcccccagc gtctggagga ttccacaccc tcgcaccgca
     1681 ggggcgagga agtgggcgga gtccggtttt ggcgccagcc gctgaggctg ccaagcagaa
     1741 aagccaccgc tgaggagact ccggtcactg tcctcgcccc gcctccccct tccctcccct
     1801 tggggaccac cgggcgccac gccgcgaacg gtaagtgccg cggtcgtcgg cgcctccgcc
     1861 ctccccctag ggccccaatt cccagcgggc gcggcgcgcg gcccctcccc ccgccgggcg
     1921 cgcgcccgct gccccgccct tcgtggccgc ccggcgtggg cggtgccacc cctccccccg
     1981 gcggccccgc gcgcagctcc cggctccctc ccccttcgga tgtggcttga gctgtaggcg
     2041 cggagggtac cagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta
     2101 tcagtgatag agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga
     2161 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
     2221 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
     2281 tcagtgatag agaacgtata agctttaggc gtgtacggtg ggcgcctata aaagcagagc
     2341 tcgtttagtg aaccgtcaga tcgcctggag caattccaca acacttttgt cttataccaa
     2401 ctttccgtac cacttcctac cctcgtaaag gatccgcccc accatggctc ctaagaaaaa
     2461 acggaaggtg tctagcgaga caggccctgt ggccgtggat cctacactgc ggagaagaat
     2521 cgagccccac gagttcgagg tgttcttcga ccccagagag ctgcggaaag agacatgcct
     2581 gctgtacgag atcaactggg gcggcagaca ctctatctgg cggcacacaa gccagaacac
     2641 caacaagcac gtggaagtga actttatcga gaagtttacg accgagcggt acttctgccc
     2701 caacaccaga tgcagcatca cctggtttct gagctggtcc ccttgcggcg agtgcagcag
     2761 agccatcacc gagtttctgt ccagatatcc ccacgtgacc ctgttcatct atatcgcccg
     2821 gctgtaccac cacgccgatc ctagaaatag acagggactg cgcgacctga tcagcagcgg
     2881 agtgaccatc cagatcatga ccgagcaaga gagcggctac tgctggcgga acttcgtgaa
     2941 ctacagcccc agcaacgaag cccactggcc tagatatcct cacctgtggg tccgactgta
     3001 cgtgctggaa ctgtactgca tcatcctggg cctgcctcca tgcctgaaca tcctgagaag
     3061 aaagcagcct cagctgacct tcttcacaat cgccctgcag agctgccact accagagact
     3121 gcctccacac atcctgtggg ccaccggact gaagtctggc tctgagacac ctggcacaag
     3181 cgagtctgcc acacctgagt ctgacaagaa atactcaatc gggctggcca tcggaactaa
     3241 ctcagtgggg tgggcagtca ttactgacga gtacaaagtg ccaagcaaga aatttaaggt
     3301 cctgggcaac accgataggc actccatcaa gaaaaatctg attggggccc tgctgttcga
     3361 ctctggagag acagctgaag caactagact gaaaaggact gctagaaggc gctatacccg
     3421 gcgaaagaat cgcatctgct acctgcagga gattttctct aacgaaatgg ccaaggtgga
     3481 cgatagtttc tttcatcggc tggaggaatc attcctggtc gaggaagata agaaacacga
     3541 gagacatcct atctttggaa acattgtgga cgaggtcgct tatcacgaaa aataccccac
     3601 catctatcat ctgcgcaaga aactggtgga ctctacagat aaagcagacc tgcggctgat
     3661 ctatctggcc ctggctcaca tgattaagtt cagaggccat tttctgatcg agggagatct
     3721 gaacccagac aatagcgatg tggacaagct gttcatccag ctggtccaga catacaatca
     3781 gctgtttgag gaaaacccta ttaatgcatc tggcgtggac gcaaaagcca tcctgagtgc
     3841 caggctgtct aagagtagaa ggctggagaa cctgatcgct cagctgccag gcgaaaagaa
     3901 aaacggcctg tttggaaatc tgattgcact gtcactggga ctgacaccta acttcaagag
     3961 caattttgat ctggccgagg acgctaaact gcagctgagc aaggacactt atgacgatga
     4021 cctggataac ctgctggctc agatcggaga tcagtacgca gacctgttcc tggccgctaa
     4081 gaatctgtct gacgctatcc tgctgagtga tattctgcgg gtgaacaccg agattacaaa
     4141 agcccctctg tcagctagca tgatcaagag atatgacgag caccatcagg atctgaccct
     4201 gctgaaggca ctggtgcgcc agcagctgcc cgagaagtac aaggaaatct tctttgatca
     4261 gagtaagaac gggtacgccg gttatattga cggcggagct tcacaggagg aattctacaa
     4321 gtttatcaaa cctattctgg agaagatgga cggcaccgag gaactgctgg tgaaactgaa
     4381 tcgcgaggac ctgctgcgca agcagcggac atttgataac ggctccatcc cccaccagat
     4441 tcatctggga gagctgcacg caatcctgcg acgacaggaa gacttctacc catttctgaa
     4501 ggataaccgc gagaagatcg aaaaaattct gaccttccgg atcccttact atgtggggcc
     4561 cctggcaagg ggtaattccc gctttgcctg gatgacacgg aaatctgagg aaacaatcac
     4621 tccttggaac ttcgaggaag tggtcgataa gggagcttcc gcacagtctt tcatcgagag
     4681 aatgacaaac ttcgacaaaa acctgccaaa tgagaaagtg ctgcctaagc acagtctgct
     4741 gtacgagtat ttcacagtct ataacgaact gactaaggtg aaatacgtca ccgaggggat
     4801 gaggaagccc gccttcctga gcggtgaaca gaagaaagct atcgtggacc tgctgtttaa
     4861 aaccaatcgc aaggtgacag tcaagcagct gaaggaggac tacttcaaga aaattgaatg
     4921 tttcgattct gtggagatca gtggcgtcga agacagattt aacgcttctc tgggaaccta
     4981 ccacgatctg ctgaagatca ttaaggataa agacttcctg gacaacgagg aaaatgagga
     5041 tatcctggaa gacattgtgc tgaccctgac actgtttgag gatcgcgaaa tgatcgagga
     5101 acggctgaaa acttatgccc atctgttcga tgacaaggtg atgaaacagc tgaagcgaag
     5161 aaggtacacc ggctggggac gactgagcag aaagctgatc aacggcattc gggacaaaca
     5221 gagtggaaag actatcctgg actttctgaa atcagatggc ttcgctaaca gaaattttat
     5281 gcagctgatt cacgatgaca gcctgacctt caaagaggat atccagaagg cacaggtgtc
     5341 cgggcagggt gactctctgc acgagcatat cgcaaacctg gccgggtccc ccgccatcaa
     5401 gaaaggtatt ctgcagaccg tgaaggtggt cgatgagctg gtgaaagtca tgggcaggca
     5461 taagccagaa aacatcgtga ttgagatggc ccgcgaaaat cagaccacac agaaaggaca
     5521 gaagaacagc cgcgagcgga tgaaaaggat cgaggaaggc attaaggaac tgggatccca
     5581 gatcctgaaa gagcaccctg tggaaaacac tcagctgcag aatgagaagc tgtatctgta
     5641 ctatctgcag aatgggcggg atatgtacgt ggaccaggag ctggatatta accgactgtc
     5701 tgattacgac gtggatcata tcgtcccaca gtcattcctg aaagatgaca gcattgacaa
     5761 taaggtgctg acccggagtg acaaaaaccg aggaaagagt gataatgtcc cttcagagga
     5821 agtggtcaag aaaatgaaga actactggag acagctgctg aatgccaaac tgatcacaca
     5881 gcgaaagttt gataacctga ctaaagctga gagagggggt ctgtcagaac tggacaaagc
     5941 aggcttcatc aagcgacagc tggtggagac cagacagatc acaaagcacg tcgctcagat
     6001 tctggatagc aggatgaaca caaagtacga tgagaatgac aaactgatcc gcgaagtgaa
     6061 ggtcattact ctgaagtcaa aacttgtgag cgacttcaga aaggatttcc agttctacaa
     6121 agtcagggag atcaacaatt atcaccatgc tcatgacgca tacctgaacg cagtggtcgg
     6181 gaccgccctg attaagaaat accccaaact ggagagcgaa ttcgtgtacg gtgactataa
     6241 ggtgtacgat gtcagaaaaa tgatcgccaa gagtgagcag gaaattggaa aagccaccgc
     6301 taagtatttc ttttactcaa acatcatgaa tttctttaag actgagatca ccctggcaaa
     6361 tggggaaatc cgaaagagac cactgattga gactaacggc gagaccggag aaatcgtgtg
     6421 ggacaagggt agggattttg ccacagtgcg caaggtcctg tccatgcctc aagtgaatat
     6481 tgtcaagaaa acagaggtgc agactggcgg attcagtaag gaatcaattc tgcccaaacg
     6541 gaactctgat aagctgatcg cccgaaagaa agactgggat cccaagaaat atgggggttt
     6601 cgactcccca acagtggctt actctgtcct ggtggtcgca aaggtggaga aggggaaaag
     6661 caagaaactg aaatccgtca aggagctgct gggtatcact attatggaga ggagctcctt
     6721 cgagaagaac cccatcgatt ttctggaggc taaaggctat aaggaagtga agaaagacct
     6781 gatcattaaa ctgccaaagt acagcctgtt tgagctggaa aacggaagga agcgaatgct
     6841 ggcatccgca ggagagctgc agaagggtaa tgaactggcc ctgccttcta agtacgtgaa
     6901 cttcctgtat ctggctagcc actacgagaa gctgaaaggc tcccccgagg ataacgaaca
     6961 gaaacagctg tttgtggagc agcacaagca ttatctggac gagatcattg aacagattag
     7021 cgagttctcc aaaagagtga tcctggctga cgcaaatctg gataaggtcc tgagcgcata
     7081 caacaaacac agagataagc caatcaggga gcaggccgaa aatatcattc atctgttcac
     7141 tctgaccaac ctgggagccc ctgcagcctt caagtatttt gacactacca tcgatcggaa
     7201 acgatacaca tccactaagg aggtgctgga cgctaccctg attcaccaga gcattaccgg
     7261 cctgtatgaa acaaggattg acctgtctca gctggggggc gactctggcg gcagcaccaa
     7321 cctgagcgac atcatcgaga aagagacagg caagcagctg gtcatccaag agtccatcct
     7381 gatgctgcct gaagaggtgg aagaagtgat cggcaacaag cccgagtccg acatcctggt
     7441 gcacaccgcc tacgatgaga gcaccgacga gaacgtgatg ctgctgacct ctgacgcccc
     7501 tgagtacaag ccttgggctc tcgtgatcca ggacagcaac ggcgagaaca agatcaagat
     7561 gctgagcggc ggcagcccta agaagaagcg gaaggtcgag gaccccaaga aaaaacggaa
     7621 ggtgtgataa aaactcgcga atcgatcgcc caccccaact tgtttattgc agcttataat
     7681 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7741 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     7801 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7861 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7921 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7981 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     8041 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     8101 gcggtatcag ctcactcaaa ggacggccgc acatgtaagg aaaattttag ggatgttaaa
     8161 gaaaaaaata acacaaaaca aaatataaaa aaaatctaac ctcaagtcaa ggcttttcta
     8221 tggaataagg aatggacagc agggggctgt ttcatatact gatgacctct ttatagccaa
     8281 cctttgttca tggcagccag catatgggca tatgttgcca aactctaaac caaatactca
     8341 ttctgatgtt ttaaatgatt tgccctccca tatgtccttc cgagtgagag acacaaaaaa
     8401 ttccaacaca ctattgcaat gaaaataaat ttcctttatt agccagaagt cagatgctca
     8461 aggggcttca tgatgtcccc ataatttttg gcagagggaa aaagatctca gtggtatttg
     8521 tgagccaggg cattggccac accagtcacc accttctgat aggcagcctg cacctgagga
     8581 gtgaattcac gcgttttagc ctggcagcat atcgagatcg aagtcatcga gagcgtcagc
     8641 agggagcata tccagatcaa aatcatccag ggcatcggcg ggcagcatgt ccaggtcgaa
     8701 gtcgtccagg gcgtcggtgg gtccgccgct ctcgcacttc agctgcttct cgaggccgca
     8761 gatgatcagt tccaggccga acaggaaggc gggctcggcg ccctgtctgt cgaacagctc
     8821 gatggcctgc ttcagcaggg ggggcatgct gtcggtggtg ggtgtctctc tttcctcttt
     8881 ggcgacctgg tgttcctgtt cttccagcac gcagcccagg gtgaagtggc ccacggcgct
     8941 cagggcgtac agggcgtttt ccaggctgaa gccctgctgg cacaggaagg ccagctggtt
     9001 ttccagtgtc tcgtactgct tctcggtggg tctggtgccc aggtgcactt tggcgccgtc
     9061 ccggtggctc agcagggcgc atctgtagct cttggcgttg ttccgcagga agtcctgcca
     9121 gctctcgcct tccagagggc agctgtgggt gtggtgccgg tccagcatct cgatgggcag
     9181 ggcgtccagc agggcccgct tgttcttcac gtgccagtac agggtgggct gttccacgcc
     9241 cagtttctgg gccagcttcc gggtggtcag gccctcgatg cccacgccgt tcagcagttc
     9301 cagggcgctg ttgatcactt tgctcttgtc cagccggctc atgtcgacat tctttgccaa
     9361 aatgatgaga cagcacaata accagcacgt tgcccaggag ctgtaggaaa aagaagaagg
     9421 catgaacatg gttagcagag gctctagagc cgccggtcac acgccagaag ccgaaccccg
     9481 ccctgccccg tcccccccga aggcagccgt ccccccgcgg acagccccga ggctggagag
     9541 ggagaagggg acggcggcgc ggcgacgcac gaaggccctc cccgcccatt tccttcctgc
     9601 cggcgccgca ccgcttcgcc ccgcgcccgc tagagggggt gcggcggcgc ctcccagatt
     9661 tcggctccgc acagatttgg gacaaaggaa gtccctgcgc cctctcgcac gattaccata
     9721 aaaggcaatg gctgcggctc gccgcgcctc gacagccgcc ggcgctccgg gggccgccgc
     9781 gccccycccc cgagcccycc ccggcccgrg gcggccccgc cccgcccggc acccccmcct
     9841 gccgccaccc cccgcccggc acggcgagcc ccgcgccacg ccccgtacgg agccccgcac
     9901 ccgaagccgg gccgtgctca gcaactcggg gaggggggtg cagggggggt tgcagcccga
     9961 ccgacgcgcc cacaccccct gctcaccccc ccacgcacac accccgcacg cagcctttgt
    10021 tcccctcgca gccccccccg caccgcgggg caccgccccc ggccgcgctc ccctcgcgca
    10081 cactgcggag cgcacaaagc cccgcgccgc gcccgcagcg ctcacagccg ccgggcagcg
    10141 cggagccgca cgcggcgctc cccacgcaca cacacacgca cgcacccccc gagccgctcc
    10201 ccccgcacaa agggccctcc cggagcccct caaggctttc acgcagccac agaaaagaaa
    10261 caagccgtca ttaaaccaag cgctaattac agcccggagg agaagggccg tcccgcccgc
    10321 tcacctgtgg gagtaacgcg gtcagtcaga gccggggcgg gcggcgcgag gcggcggcgg
    10381 agcggggcac ggggcgaagg cagcgcgcag cgactcccgc ccgccgcgcg cttcgctttt
    10441 tatagggccg ccgccgccgc cgcctcgcca taaaaggaaa ctttcggagc gcgccgctct
    10501 gattggctgc cgccscmccy cyccsccycs ccccsccccs ccccycsccc csccccsccc
    10561 csccysgcgc scsccccccc cccccccccg cccccatcgc tgcacaaaat aattaaaaaa
    10621 taaataaata caaaattggg ggtggggagg ggggggagat ggggagagtg aagcagaacg
    10681 tggggctcac ctcgaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
    10741 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
    10801 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
    10861 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
    10921 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
    10981 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
    11041 ttgattacta ttaataacta gtcaataatc aatgtcgaga cccatagagc ccaccgcatc
    11101 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
    11161 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
    11221 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
    11281 agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttata acttcgtata
    11341 atgtatgcta tacgaagtta tggtacctta ctcgtccatg ccgccggtgg agtggcggcc
    11401 ctcggcgcgt tcgtactgtt ccacgatggt gttacttgta cagctcgtcc attccgcctg
    11461 tgctgtgtct gccttcggct ctctcgtact gttccacgat ggtgtagtcc tcgttgtggg
    11521 acacgatgtc cagcttgata tccacgatgt aggcgccagg cagctgcacg ggtttcttgg
    11581 ccttgtaggt ggttttcact tcggcggcat agtggccgcc atccttcagc ttcagccgct
    11641 tcttgatctc ggacttcagg gcgccatcct cagggtacat tctttcctcg ctggcttccc
    11701 agcccatggt ctttttctgc atcacggggc catcagaggg aaagttggtg ccccgcagct
    11761 tcactttgta gatgaacacg ccgtcctgca gagagctatc ctggttcacg tggatgatgc
    11821 cgccgtcctc gaagttcatc actctctccc atctgaagcc ctcggggaag ctcagcttga
    11881 agtagtcggg aatgtcggcg gggtgcttga tgtacacctt gctgccgtac ataaactgag
    11941 ggctcagaat gtcccaggca aaaggcagag ggccgccttt ggtcactttc agcttggcgg
    12001 tctgaaaagc ctcgtagggt ctgccctcgc cttcgccttc aatctcgaac tcgtggccgt
    12061 tcacgctgcc ttccatgtgc accttgaacc gcatgaattc tttgatgatg gccatgttgt
    12121 tctcttcgcc cttggacacc atggtggcga ccggtagcgt ggcgaccggg gtggcggtgg
    12181 ctccccctga ccctccatcc aacttaacgc gctttgcagc aggacctcca gaaccgccgt
    12241 cgagtttgac ccgtttagcg gcagggaccg gtgggccagg attctcctcg acgtcaccgc
    12301 atgttagcag acttcctctg ccctctcccg atcccgatcc atttcgggta tatttgagtg
    12361 gaatgagttc ttcaatcgta gttttgacta acttgccatt catttctatt aacacaaaac
    12421 aatctggtgc atagtctgaa atcaactccc tacacatacc acaaggactt accactcgaa
    12481 tacttctatc tacttcgtca gaataagggt gtctaacagc tacaatcgtg tcaaaatcct
    12541 tttgtccatt cgaaactgca ctaccaatcg caatggcttc tgcacaaaca gttactcgtc
    12601 ctatatacgc ttcaatatgt actgccgaaa tgatttctcc tgttttcgta cgaattgccg
    12661 ctcccacatg atgtttatta tcctcataaa gcattgtaat cttctctgtc gctacttcta
    12721 ctaattctag atcctgctgg gagatgttga aggtcttcat ggtggcgcta gcgtaggcgc
    12781 cggtcacagc ttggatctgt aacggcgcag aacagaaaac gaaacaaaga cgtagagttg
    12841 agcaagcagg gtcaggcaaa gcgtggagag ccggctgagt ctaggtaggc tccaagggag
    12901 cgccggacaa aggcccggtc tcgacctgag ctttaaactt acctagacgg cggacgcagt
    12961 tcaggaggca ccacaggcgg gaggcggcag aacgcgactc aaccggcgtg gatggcggcc
    13021 tcaggtaggg cggcgggcgc gtgaaggaga gatgcgagcc cctcgaagct tcagctgtgt
    13081 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
    13141 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
    13201 cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca acttctcggg
    13261 gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagcgatcgc agatcatgca
    13321 tccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg
    13381 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
    13441 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
    13501 acgggggagg ggcaaacaac agatggctgg caactagaag gcacaggttt ttagaagaac
    13561 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
    13621 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
    13681 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
    13741 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
    13801 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
    13861 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
    13921 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
    13981 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
    14041 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
    14101 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
    14161 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
    14221 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
    14281 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
    14341 atcctagggc cgggattctc ctccacgtca ccgcatgtta gaagacttcc tctgccctct
    14401 ccgctgccag atctctcgag gccctgtggg aggaagagaa gaggtcagat aacttcgtat
    14461 aatgtatgct atacgaagtt atgcggccgc tgggagaagg gtaagagagc tcaatcctca
    14521 tcttacatag aaggaagtca gtagataata tcttacattt aaaagtcaag aaataacagt
    14581 ctgaccatac tatctagaaa tatggaagtt aacagtaagg aagttaaaac agttgaaagt
    14641 agtttcgtct agaaagaaag acttggggaa gaacaaagaa ggctaatttt cattatgagc
    14701 tttattgtaa cagaaaagag accactaact cagctaggtt ttcccaattc ctcaacactt
    14761 tggggcacag tgtttagtca gagctgtgat cactctctga gcacatggat tgtatcttaa
    14821 ttttcatctc attgaaaata tgtaccattt gtccaagtgg aaactgggct gaagcatgac
    14881 tgtgaccacc accaaagaaa tggctagact cgctaaggtc tgctggggtg acccgtgatt
    14941 gtgttgtgct tgatcttgtt gagcatggct cttttaagtc accatacctg cccctgccaa
    15001 tgcaccaaac accaacttaa agggactttc agcttttgta agtcagtctc agccttactt
    15061 ctgaaccatt tctttgactt ctttcatgac tctgatcctg tcccagtttt aagcctccca
    15121 gccaccagtc atgctgtttt gctctgctat ttacattctt ctctacctac tttggacttg
    15181 atggtatgcc tactatatta gttatttatt gctgtgtaac aaaccactct gaaacatagt
    15241 ggcttacaac cacaaccatt ttattttatt atgaaactgt gggtcagaat gtatattggc
    15301 ccattggcat gattctcctg ctattgaaaa aggaagagta tgagtattca acatttccgt
    15361 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
    15421 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
    15481 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
    15541 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
    15601 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
    15661 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
    15721 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
    15781 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
    15841 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
    15901 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
    15961 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
    16021 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
    16081 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    16141 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
    16201 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
    16261 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
    16321 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    16381 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    16441 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    16501 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    16561 gtagcaccgc ctacatacct cgctctgcta atcc
//
