LOCUS       sequence-468067-       13907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cytosine Base Editor for Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13907)
  AUTHORS   Ng EFY, Meitinger F
  TITLE     Genetic Engineering and Screening Using Base Editing and Inducible 
            Gene Knockout.
  JOURNAL   Methods Mol Biol. 2025;2872:167-187. doi: 
            10.1007/978-1-0716-4224-5_12.
  PUBMED    39616576
REFERENCE   2  (bases 1 to 13907)
  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"; journalName: "Methods Mol
            Biol."; date: "2025"; volume: "2872:167-187. doi"; pages: " 
            10.1007/978-1-0716-4224-5_12"
COMMENT     Sequence Label: pMO819
FEATURES             Location/Qualifiers
     source          1..13907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    216..333
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(631..675)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    860..1093
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1586..1711
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1758..1938
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1956..2154)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1981..2001)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2156..2535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2707..2726
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2801..2905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2906..2974,2975..3766)
                     /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(3124..3143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3937..4525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4426..4445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4679..4696
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4813..4834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4849..4854,4855..4872,4873..4879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4887..4903
                     /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)."
     primer_bind     4892..4914
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4911..4927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4911..4927
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4976..5097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4987..5006)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5041..5060
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5227..5601)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTKIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(5620..5667)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      5729..5863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5783..5802)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6008..6028
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6037..6465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6147..6168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6359..6378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6511..6735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6724..6741
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6767..6947
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     7452..7468
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7453..7469
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7472..8060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7578..7589
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7987..8007
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8085..8483)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(8484..8540)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(8550..8570)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8583..8831)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(join(8844..12917,12918..12920,12921..12944))
                     /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"
     CDS             complement(8844..12944)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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"
     CDS             complement(12993..13679)
                     /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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             complement(13704..13724)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(13731..13796)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      13796..13805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(13819..13907,1..123))
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt ctctaggcac
       61 cggatcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat gtgcgctctg
      121 cccactgacg ggcaccggag ccaattccca ctcctttcaa gacctagcta gctaattaac
      181 caaactggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
      241 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
      301 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtctg
      361 cagaattggc gcacgcagtg ccgatccgtt cactaatcga atggatctgt ctctgtctct
      421 ctctccacct tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg
      481 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
      541 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
      601 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
      661 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
      721 tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc
      781 cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc
      841 agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat
      901 tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc
      961 aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg
     1021 taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag
     1081 gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt
     1141 ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata attcacttct
     1201 ccaattgtcc ctcatatctc ctcctccagg tctgaagatc agcggccgct tgctgtgcgg
     1261 tggtcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta
     1321 tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt
     1381 cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg
     1441 atgtttctaa caggccagga ttaactgcga atcgttctag ctccctgctt gcccatacta
     1501 tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg
     1561 atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc
     1621 cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg
     1681 cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt
     1741 ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct
     1801 ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct
     1861 ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg
     1921 gtctaaccag agagacccag tacaggcaaa acgcgctgct tatatagacc tcccaccgta
     1981 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     2041 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     2101 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     2161 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     2221 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     2281 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     2341 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     2401 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     2461 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     2521 tcaataatca atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc
     2581 taaccctaag cagattcttc atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa
     2641 ttttgctcgc gcactactca gcgacctcca acacacaagc agggagcaga tactggctta
     2701 actatgcggc atcagagcag attgtactga gagtgcacca taggggatcg ggagatctcc
     2761 cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2821 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2881 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2941 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3001 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3061 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3121 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3181 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3241 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3301 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3361 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3421 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3481 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3541 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3601 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3661 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3721 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3781 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3841 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3901 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttttgcgcgt
     3961 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4021 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4081 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4141 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4201 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4261 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4321 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4381 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4441 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4501 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4561 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4621 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4681 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4741 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4801 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     4861 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4921 ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga catgataaga
     4981 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     5041 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttggg
     5101 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     5161 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtag
     5221 cacgtgtcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg ggtcgcgcag
     5281 ggcgaactcc cgcccccacg gctgctcgcc gatcttggtc atggccggcc cggaggcgtc
     5341 ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca ggccgcgcac
     5401 ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc tgatgaacag
     5461 ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc gggagaaccc
     5521 gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga gcaccggaac
     5581 ggcactggtc aacttggcca tggtttagtt cctcaccttg tcgtattata ctatgccgat
     5641 atactatgcc gatgattaat tgtcaacacg tgctgatcag atccgaaaat ggatatacaa
     5701 gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5761 aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     5821 tggggcggag aatgggcgga actgggcgga gttaggggcg ggactatggt tgctgactaa
     5881 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     5941 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     6001 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
     6061 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     6121 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     6181 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6241 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6301 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6361 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6421 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag
     6481 agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc
     6541 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     6601 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     6661 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     6721 aactagaagg cacagtcgag gctgatcagc gggtttaaac gggccctgct agagattttc
     6781 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     6841 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     6901 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acagtccgga
     6961 tgcagctctc gggccatgtg atgaaatgct aggcggctgt caaacctcca ctctaatact
     7021 tctctctccg ggtcatccat cccatgcagg ctcacagggt gtaacaagcg ggtgttctct
     7081 ccttcattgg cttcttctac cttctcttgc tcaactggta ctagcttgta gcaccatcca
     7141 aaggtcagtg gatatctgat ccctggccct ggtgtgtagt tctgccaatc agggaagtag
     7201 ccttgtgtgt ggtagatcca cagatcaagg atatcttgtc ttcgttggga gtgaattagc
     7261 ccttccagtc cccccttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc
     7321 attggtctta aaggtacctg aggtgtgact ggaaaaccca cctcctcctc ctcttgtgct
     7381 tctagccagg cacaatcagc attggtagct gctgtattgc tacttgtgat tgctccatgt
     7441 ttttctaggt ctcgaggtcg acggtatcga tgcggggagg cggcccaaag ggagatccga
     7501 ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag caggccgcgg
     7561 gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt cccgcgcaga
     7621 atccaggtgg caacacaggc gagcagccaa ggaaaggacg atgatttccc cgacaacacc
     7681 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     7741 atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag gagctgacag
     7801 gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca caccacgcca
     7861 cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga tttatacaag
     7921 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     7981 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     8041 ttgtaatcca gaggttgatt accgataagc ttgatatcga attcttagcc ctcccacaca
     8101 taaccagagg gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc
     8161 actatggctt tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg
     8221 ctcaagatgc ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc
     8281 gcagcagcag cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat
     8341 acattgacac cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag
     8401 tcttcagaga tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct
     8461 tcttgagaca aaggcttggc catcggtcca ggattctctt cgacatctcc ggcttgtttc
     8521 agcagagaga agtttgttgc gccggatccg actttcctct tcttcttggg agaaccacca
     8581 gagagcatct taatcttgtt ctcaccgttg ctatcctgta tgaccagagc ccaaggcttg
     8641 tattcagggg cgtcgctagt cagaagcatg acattctcgt cggtgctctc gtcgtaggcg
     8701 gtgtgcacga gtatatcgct ttccggcttg ttcccaatga cttcttccac ctcctctggg
     8761 agcatgagga tggattcctg gataaccagt tgcttaccgg tctccttttc aataatatct
     8821 gacagattag tagatccgcc tgaatcgcct cccagctgag acaggtcgat ccgtgtctcg
     8881 tacaggccgg tgatgctctg gtggatcagg gtggcgtcca gcacctcttt ggtgctggtg
     8941 tacctcttcc ggtcgatggt ggtgtcaaag tacttgaagg cggcaggggc tcccagattg
     9001 gtcagggtaa acaggtggat gatattctcg gcctgctctc tgatgggctt atcccggtgc
     9061 ttgttgtagg cggacagcac tttgtccaga ttagcgtcgg ccaggatcac tctcttggag
     9121 aactcgctga tctgctcgat gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc
     9181 tgtttctgct cattatcctc gggggagccc ttcagcttct catagtggct ggccaggtac
     9241 aggaagttca catatttgga gggcagggcc agttcgtttc ccttctgcag ttcgccggca
     9301 gaggccagca ttctcttccg gccgttttcc agctcgaaca gggagtactt aggcagcttg
     9361 atgatcaggt cctttttcac ttctttgtag cccttggctt ccagaaagtc gatgggattc
     9421 ttctcgaagc tgcttctttc catgatggtg atccccagca gctctttcac actcttcagt
     9481 ttcttggact tgcccttttc cactttggcc accaccagca cagaataggc cacggtgggg
     9541 ctgtcgaagc cgccgtactt cttagggtcc cagtccttct ttctggcgat cagcttatcg
     9601 ctgttcctct tgggcaggat agactctttg ctgaagccgc ctgtctgcac ctcggtcttt
     9661 ttcacgatat tcacttgggg catgctcagc actttccgca cggtggcaaa atcccggccc
     9721 ttatcccaca cgatctcccc ggtttcgccg tttgtctcga tcagaggccg cttccggatc
     9781 tcgccgttgg ccagggtaat ctcggtcttg aaaaagttca tgatgttgct gtagaagaag
     9841 tacttggcgg tagccttgcc gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg
     9901 tacaccttgt agtcgccgta cacgaactcg ctttccagct tagggtactt tttgatcagg
     9961 gcggttccca cgacggcgtt caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg
    10021 cgcactttgt aaaactggaa atccttccgg aaatcggaca ccagcttgga cttcagggtg
    10081 atcactttca cttcccggat cagcttgtca ttctcgtcgt acttagtgtt catccgggag
    10141 tccaggatct gtgccacgtg ctttgtgatc tgccgggttt ccaccagctg tctcttgatg
    10201 aagccggcct tatccagttc gctcaggccg cctctctcgg ccttggtcag attgtcgaac
    10261 tttctctggg taatcagctt ggcgttcagc agctgccgcc agtagttctt catcttcttc
    10321 acgacctctt cggagggcac gttgtcgctc ttgccccggt tcttgtcgct tctggtcagc
    10381 accttgttgt cgatggagtc gtccttcaga aagctctgag gcacgatatg gtccacatcg
    10441 tagtcggaca gccggttgat gtccagttcc tggtccacgt acatatcccg cccattctgc
    10501 aggtagtaca ggtacagctt ctcgttctgc agctgggtgt tttccactgg gtgttctttc
    10561 aggatctggc tgcccagctc tttgatgccc tcttcgatcc gcttcattct ctcgcggctg
    10621 ttcttctgtc ccttctgggt ggtctggttc tctctggcca tttcgatcac gatgttctcg
    10681 ggcttgtgcc ggcccatcac tttcacgagc tcgtccacca ccttcactgt ctgcaggatg
    10741 cccttcttaa tggcggggct gccggccaga ttggcaatgt gctcgtgcag gctatcgccc
    10801 tggccggaca cctgggcttt ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc
    10861 agctgcatga agtttctgtt ggcgaagccg tcggacttca ggaaatccag gattgtcttg
    10921 ccggactgct tgtcccggat gccgttgatc agcttccggc tcagcctgcc ccagccggtg
    10981 tatctccgcc gcttcagctg cttcatcact ttgtcgtcga acaggtgggc ataggttttc
    11041 agccgttcct cgatcatctc tctgtcctca aacagtgtca gggtcagcac gatatcttcc
    11101 agaatgtcct cgttttcctc attgtccagg aagtccttgt ccttgataat tttcagcaga
    11161 tcgtggtatg tgcccaggga ggcgttgaac cgatcttcca cgccggagat ttccacggag
    11221 tcgaagcact cgattttctt gaagtagtcc tctttcagct gcttcacggt cactttccgg
    11281 ttggtcttga acagcaggtc cacgatggcc tttttctgct cgccgctcag gaaggcgggc
    11341 tttctcattc cctcggtcac gtatttcact ttggtcagct cgttatacac ggtgaagtac
    11401 tcgtacagca ggctgtgctt gggcagcacc ttctcgttgg gcaggttctt atcgaagttg
    11461 gtcatccgct cgatgaagct ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc
    11521 cagggggtga tggtttcctc gctctttctg gtcatccagg cgaatctgct gtttcccctg
    11581 gccagagggc ccacgtagta ggggatgcgg aaggtcagga tcttctcgat cttttcccgg
    11641 ttgtccttca ggaatgggta aaaatcttcc tgccgccgca gaatggcgtg cagctctccc
    11701 aggtggatct ggtgggggat gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc
    11761 tctctgttca gcttcacgag cagttcctcg gtgccgtcca tcttttccag gatgggcttg
    11821 atgaacttgt agaactcttc ctggctggct ccgccgtcaa tgtagccggc gtagccgttc
    11881 ttgctctggt cgaagaaaat ctctttgtac ttctcaggca gctgctgccg cacgagagct
    11941 ttcagcaggg tcaggtcctg gtggtgctcg tcgtatctct tgatcataga ggcgctcagg
    12001 ggggccttgg tgatctcggt gttcactctc aggatgtcgc tcagcaggat ggcgtcggac
    12061 aggttcttgg cggccagaaa caggtcggcg tactggtcgc cgatctgggc cagcaggttg
    12121 tccaggtcgt cgtcgtaggt gtccttgctc agctgcagtt tggcatcctc ggccaggtcg
    12181 aagttgctct tgaagttggg ggtcaggccc aggctcaggg caatcaggtt gccgaacagg
    12241 ccattcttct tctcgccggg cagctgggcg atcagatttt ccagccgtct gctcttgctc
    12301 agtctggcag acaggatggc cttggcgtcc acgccgctgg cgttgatggg gttttcctcg
    12361 aacagctggt tgtaggtctg caccagctgg atgaacagct tgtccacgtc gctgttgtcg
    12421 gggttcaggt cgccctcgat caggaagtgg ccccggaact tgatcatgtg ggccagggcc
    12481 agatagatca gccgcaggtc ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg
    12541 tagatggtgg ggtacttctc gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg
    12601 tgccgctcgt gcttcttatc ctcttccacc aggaaggact cttccagtct gtggaagaag
    12661 ctgtcgtcca ccttggccat ctcgttgctg aagatctctt gcagatagca gatccggttc
    12721 ttccgtctgg tgtatcttct tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg
    12781 ccgctgtcga acagcagggc tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg
    12841 cccagcacct tgaatttctt gctgggcacc ttgtactcgt cggtgatcac ggcccagccc
    12901 acagagttgg tgccgatggc caggccgatg ctgtacttct tgtcactttc gggtgtggcg
    12961 gactctgagg tcccgggagt ctcgctgccg cttttcaacc cggtggccca gagaatgtgt
    13021 gggggcagtc gctggtaatg acaagactga agagcgatgg taaagaatgt cagctgtggc
    13081 tgcttccttc tcagaatgtt gagacaagga ggcaggccca gtatgatgca gtacagttca
    13141 agaacgtaca gtcgtaccca cagatgggga tacctaggcc agtgggcttc attactcggg
    13201 ctataattca caaagtttct ccagcagtat cctgactcct gctcagtcat aatttggata
    13261 gtcacacctg aagagatcaa atcccgcagg ccttgtcgat tgcgggggtc agcgtggtgg
    13321 tacagccttg cgatgtaaat aaacagagtg acgtggggat accttgacag gaattcagtg
    13381 atggccctac tacattcgcc gcatgggctc cagctgagaa accaggtaat gctgcacctt
    13441 gtgttcggac agaaatatct ttctgtcgtg aacttctcga tgaagttgac ttcgacgtgc
    13501 ttgttagtgt tctgtgatgt atgtcgccaa atggagtgcc ggccccccca attaatttcg
    13561 taaagcaggc aggtctcctt gcggagctct ctcggatcga agaatacctc aaactcatgg
    13621 ggctcgatcc gccgtctcaa tgtggggtcc acagccactg ggccagtctc tgagctcatg
    13681 gctgctggga ctccgtggat accgaccttc cgcttcttct ttggggccat cttatcgtca
    13741 tcgtctttgt aatcaatatc atgatccttg tagtctccgt cgtggtcctt atagtccatg
    13801 gtggctctag aaccggtcct gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac
    13861 ggcgactact gcacttatat acggttctcc cccaccctcg ggaaaaa
//
