LOCUS       sequence-320698-        9849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian CG-to-GC base editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9849)
  AUTHORS   Koblan LW, Arbab M, Shen MW, Hussmann JA, Anzalone AV, Doman JL, 
            Newby GA, Yang D, Mok B, Replogle JM, Xu A, Sisley TA, Weissman JS, 
            Adamson B, Liu DR
  TITLE     Efficient C*G-to-G*C base editors developed using CRISPRi screens, 
            target-library analysis, and machine learning.
  JOURNAL   Nat Biotechnol. 2021 Jun 28. pii: 10.1038/s41587-021-00938-z. doi: 
            10.1038/s41587-021-00938-z.
  PUBMED    34183861
REFERENCE   2  (bases 1 to 9849)
  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: "Nat 
            Biotechnol. 2021 Jun 28. pii: 10.1038/s41587-021-00938-z. doi: 
            10.1038/s41587-021-00938-z."
COMMENT     Sequence Label: pCMV_UdgX-APOBEC1-UdgX-nCas9
FEATURES             Location/Qualifiers
     source          1..9849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1189..1872
                     /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(2692..2715,2716..2718,2719..6792)
                     /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             2692..6792
                     /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             6835..6855
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6864..6881
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6904..6921)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6910..7134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7205..7221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7205..7221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7218..7240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7229..7245
                     /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(7253..7259,7260..7277,7278..7283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7298..7319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7436..7453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7607..8195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7687..7706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8366..9157,9158..9226))
                     /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     8989..9008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9227..9331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9410..9429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9601..9849,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtcatggc cggagcacag
      481 gattttgtcc cacatactgc cgatctggct gagctggctg ccgccgctgg agagtgtaga
      541 ggatgcgggc tgtaccggga tgccacacag gccgtgttcg gagcaggcgg caggagcgcc
      601 cgcatcatga tgatcggcga gcagccaggc gataaggagg acctggccgg cctgccattt
      661 gtgggaccag caggccggct gctggacaga gccctggagg ccgccgacat cgatagggac
      721 gccctgtacg tgaccaacgc cgtgaagcac ttcaagttta cacgcgccgc aggaggcaag
      781 aggagaatcc acaagacccc ctctcggaca gaggtggtgg cctgcagacc ttggctgatc
      841 gccgagatga ccagcgtgga gccagatgtg gtggtgctgc tgggagcaac agcagcaaag
      901 gccctgctgg gcaatgactt cagggtgacc cagcacaggg gagaggtgct gcacgtggac
      961 gatgtgccag gcgatcctgc cctggtggca accgtgcacc ctagctccct gctgagaggc
     1021 ccaaaggagg agcgggaatc cgccttcgct gggctggtgg acgacctgag agtggctgcc
     1081 gatgtgagac cttctggagg atctagcgga ggctcctctg gcagcgagac accaggaaca
     1141 agcgagtcag caacaccaga gagcagtggc ggcagcagcg gggggtcatc ctcagagact
     1201 gggcctgtcg ccgtcgatcc aaccctgcgc cgccggattg aacctcacga gtttgaagtg
     1261 ttctttgacc cccgggagct gagaaaggag acatgcctgc tgtacgagat caactgggga
     1321 ggcaggcact ccatctggag gcacacctct cagaacacaa ataagcacgt ggaggtgaac
     1381 ttcatcgaga agtttaccac agagcggtac ttctgcccca ataccagatg tagcatcaca
     1441 tggtttctga gctggtcccc ttgcggagag tgtagcaggg ccatcaccga gttcctgtcc
     1501 agatatccac acgtgacact gtttatctac atcgccaggc tgtatcacca cgcagaccca
     1561 aggaataggc agggcctgcg cgatctgatc agctccggcg tgaccatcca gatcatgaca
     1621 gagcaggagt ccggctactg ctggcggaac ttcgtgaatt attctcctag caacgaggcc
     1681 cactggccta ggtacccaca cctgtgggtg cgcctgtacg tgctggagct gtattgcatc
     1741 atcctgggcc tgcccccttg tctgaatatc ctgcggagaa agcagcccca gctgaccttc
     1801 tttacaatcg ccctgcagtc ttgtcactat cagaggctgc caccccacat cctgtgggcc
     1861 acaggcctga agtctggagg atctagcgga ggatcctctg gcagcgagac accaggaaca
     1921 agcgagtcag caacaccaga gagcagtggc ggcagcagcg gcggcagcat ggccggagca
     1981 caggattttg tcccacatac tgccgatctg gctgagctgg ctgccgccgc tggagagtgt
     2041 agaggatgcg ggctgtaccg ggatgccaca caggccgtgt tcggagcagg cggcaggagc
     2101 gcccgcatca tgatgatcgg cgagcagcca ggcgataagg aggacctggc cggcctgcca
     2161 tttgtgggac cagcaggccg gctgctggac agagccctgg aggccgccga catcgatagg
     2221 gacgccctgt acgtgaccaa cgccgtgaag cacttcaagt ttacacgcgc cgcaggaggc
     2281 aagaggagaa tccacaagac cccctctcgg acagaggtgg tggcctgcag accttggctg
     2341 atcgccgaga tgaccagcgt ggagccagat gtggtggtgc tgctgggagc aacagcagca
     2401 aaggccctgc tgggcaatga cttcagggtg acccagcaca ggggagaggt gctgcacgtg
     2461 gacgatgtgc caggcgatcc tgccctggtg gcaaccgtgc accctagctc cctgctgaga
     2521 ggcccaaagg aggagcggga atccgccttc gctgggctgg tggacgacct gagagtggct
     2581 gccgatgtga gacctagcgg tggtagttct ggcggtagct caggatctga aactcccggc
     2641 acatctgaaa gcgcgactcc cgaatcatct ggtggatctt caggtggaag tgacaagaag
     2701 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     2761 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     2821 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
     2881 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     2941 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     3001 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     3061 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     3121 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     3181 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     3241 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     3301 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     3361 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     3421 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     3481 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     3541 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     3601 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     3661 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     3721 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     3781 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     3841 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     3901 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     3961 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     4021 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     4081 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     4141 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     4201 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     4261 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     4321 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     4381 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     4441 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     4501 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     4561 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     4621 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     4681 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     4741 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     4801 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     4861 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     4921 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     4981 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     5041 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     5101 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     5161 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     5221 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     5281 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     5341 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     5401 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     5461 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     5521 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     5581 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     5641 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     5701 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     5761 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     5821 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     5881 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     5941 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     6001 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     6061 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     6121 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     6181 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     6241 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     6301 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     6361 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     6421 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     6481 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     6541 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     6601 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     6661 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     6721 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     6781 ctgggaggtg actctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagcccaag
     6841 aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     6901 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6961 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7021 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7081 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     7141 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     7201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7261 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     7321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7381 gaatcggcca acgcgcggga agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8221 ggtctgacac tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9361 tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct
     9421 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9481 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9541 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     9601 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     9661 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     9721 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     9781 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     9841 aagtgtatc
//
