LOCUS       sequence-320371-       11340 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 11340)
  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 11340)
  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_POLD2-APOBEC1-UdgX-nCas9-UdgX
FEATURES             Location/Qualifiers
     source          1..11340
                     /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             1969..2652
                     /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(3472..3495,3496..3498,3499..7572)
                     /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             3472..7572
                     /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             8326..8346
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8355..8372
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8395..8412)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8401..8625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8696..8712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8696..8712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8709..8731)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8720..8736
                     /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(8744..8750,8751..8768,8769..8774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8789..8810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8927..8944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9098..9686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9178..9197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9857..10648,10649..10717))
                     /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     10480..10499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10718..10822)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10901..10920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(11092..11340,1..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 aagtcatgtt ttctgagcag
      481 gctgcccaga gggcccacac tctactgtcc ccaccatcag ccaacaatgc cacctttgcc
      541 cgggtgccag tggcaaccta caccaactcc tcacaaccct tccggctagg agagcgcagc
      601 tttagccggc agtatgccca catttatgcc acccgcctca tccaaatgag acccttcctg
      661 gagaaccggg cccagcagca ctggggcagt ggagtgggag tgaagaagct gtgtgaactg
      721 cagcctgagg agaagtgctg tgtggtgggc actctgttca aggccatgcc gctgcagccc
      781 tccatcctgc gggaggtcag cgaggagcac aacctgctcc cccagcctcc tcggagtaaa
      841 tacatacacc cagatgacga gctggtcttg gaagatgaac tgcagcgtat caaactaaaa
      901 ggcaccattg acgtgtcaaa gctggttacg gggactgtcc tggctgtgtt tggctccgtg
      961 agagacgacg ggaagtttct ggtggaggac tattgctttg ctgaccttgc tccccagaag
     1021 cccgcacccc cacttgacac agataggttt gtgctactgg tgtccggcct gggcctgggt
     1081 ggcggtggag gcgagagcct gctgggcacc cagctgctgg tggatgtggt gacggggcag
     1141 cttggggacg aaggggagca gtgcagcgcc gcccacgtct cccgggttat cctcgctggc
     1201 aacctcctca gccacagcac ccagagcagg gattctatca ataaggccaa atacctcacc
     1261 aagaaaaccc aggcagccag cgtggaggct gttaagatgc tggatgagat cctcctgcag
     1321 ctgagcgcct cagtgcccgt ggacgtgatg ccaggcgagt ttgatcccac caattacacg
     1381 ctcccccagc agcccctcca cccctgcatg ttcccgctgg ccactgccta ctccacgctc
     1441 cagctggtca ccaaccccta ccaggccacc attgatggag tcagattttt ggggacatca
     1501 ggacagaacg tgagtgacat tttccgatac agcagcatgg aggatcactt ggagatcctg
     1561 gagtggaccc tgcgggtccg tcacatcagc cccacagccc ctgacactct aggttgttac
     1621 cccttctaca aaactgaccc gttcatcttc ccagagtgcc cgcatgtcta cttttgtggc
     1681 aacaccccca gctttggctc caaaatcatc cgaggtcctg aggaccagac agtgctgttg
     1741 gtgactgtcc ctgacttcag tgccacgcag accgcctgcc ttgtgaacct gcgcagcctg
     1801 gcctgccagc ccatcagctt ctcgggcttc ggggcagagg acgatgacct gggaggcctg
     1861 gggctgggcc cctctggagg atctagcgga ggctcctctg gcagcgagac accaggaaca
     1921 agcgagtcag caacaccaga gagcagtggc ggcagcagcg gggggtcatc ctcagagact
     1981 gggcctgtcg ccgtcgatcc aaccctgcgc cgccggattg aacctcacga gtttgaagtg
     2041 ttctttgacc cccgggagct gagaaaggag acatgcctgc tgtacgagat caactgggga
     2101 ggcaggcact ccatctggag gcacacctct cagaacacaa ataagcacgt ggaggtgaac
     2161 ttcatcgaga agtttaccac agagcggtac ttctgcccca ataccagatg tagcatcaca
     2221 tggtttctga gctggtcccc ttgcggagag tgtagcaggg ccatcaccga gttcctgtcc
     2281 agatatccac acgtgacact gtttatctac atcgccaggc tgtatcacca cgcagaccca
     2341 aggaataggc agggcctgcg cgatctgatc agctccggcg tgaccatcca gatcatgaca
     2401 gagcaggagt ccggctactg ctggcggaac ttcgtgaatt attctcctag caacgaggcc
     2461 cactggccta ggtacccaca cctgtgggtg cgcctgtacg tgctggagct gtattgcatc
     2521 atcctgggcc tgcccccttg tctgaatatc ctgcggagaa agcagcccca gctgaccttc
     2581 tttacaatcg ccctgcagtc ttgtcactat cagaggctgc caccccacat cctgtgggcc
     2641 acaggcctga agtctggagg atctagcgga ggatcctctg gcagcgagac accaggaaca
     2701 agcgagtcag caacaccaga gagcagtggc ggcagcagcg gcggcagcat ggccggagca
     2761 caggattttg tcccacatac tgccgatctg gctgagctgg ctgccgccgc tggagagtgt
     2821 agaggatgcg ggctgtaccg ggatgccaca caggccgtgt tcggagcagg cggcaggagc
     2881 gcccgcatca tgatgatcgg cgagcagcca ggcgataagg aggacctggc cggcctgcca
     2941 tttgtgggac cagcaggccg gctgctggac agagccctgg aggccgccga catcgatagg
     3001 gacgccctgt acgtgaccaa cgccgtgaag cacttcaagt ttacacgcgc cgcaggaggc
     3061 aagaggagaa tccacaagac cccctctcgg acagaggtgg tggcctgcag accttggctg
     3121 atcgccgaga tgaccagcgt ggagccagat gtggtggtgc tgctgggagc aacagcagca
     3181 aaggccctgc tgggcaatga cttcagggtg acccagcaca ggggagaggt gctgcacgtg
     3241 gacgatgtgc caggcgatcc tgccctggtg gcaaccgtgc accctagctc cctgctgaga
     3301 ggcccaaagg aggagcggga atccgccttc gctgggctgg tggacgacct gagagtggct
     3361 gccgatgtga gacctagcgg tggtagttct ggcggtagct caggatctga aactcccggc
     3421 acatctgaaa gcgcgactcc cgaatcatct ggtggatctt caggtggaag tgacaagaag
     3481 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     3541 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     3601 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
     3661 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     3721 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     3781 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     3841 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     3901 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     3961 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     4021 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     4081 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     4141 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     4201 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     4261 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     4321 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     4381 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     4441 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     4501 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     4561 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     4621 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     4681 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     4741 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     4801 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     4861 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     4921 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     4981 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     5041 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     5101 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     5161 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     5221 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     5281 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     5341 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     5401 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     5461 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     5521 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     5581 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     5641 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     5701 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     5761 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     5821 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     5881 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     5941 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     6001 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     6061 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     6121 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     6181 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     6241 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     6301 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     6361 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     6421 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     6481 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     6541 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     6601 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     6661 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     6721 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     6781 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     6841 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     6901 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     6961 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     7021 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     7081 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     7141 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     7201 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     7261 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     7321 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     7381 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     7441 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     7501 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     7561 ctgggaggtg actctggagg atctagcgga ggatcctctg gcagcgagac accaggaaca
     7621 agcgagtcag caacaccaga gagcagcggc ggcagcagcg gcggcagcat ggccggagca
     7681 caggattttg tcccacatac tgccgatctg gctgagctgg ctgccgccgc tggagagtgt
     7741 agaggatgcg ggctgtaccg ggatgccaca caggccgtgt tcggagcagg cggcaggagc
     7801 gcccgcatca tgatgatcgg cgagcagcca ggcgataagg aggacctggc cggcctgcca
     7861 tttgtgggac cagcaggccg gctgctggac agagccctgg aggccgccga catcgatagg
     7921 gacgccctgt acgtgaccaa cgccgtgaag cacttcaagt ttacacgcgc cgcaggaggc
     7981 aagaggagaa tccacaagac cccctctcgg acagaggtgg tggcctgcag accttggctg
     8041 atcgccgaga tgaccagcgt ggagccagat gtggtggtgc tgctgggagc aacagcagca
     8101 aaggccctgc tgggcaatga cttcagggtg acccagcaca ggggagaggt gctgcacgtg
     8161 gacgatgtgc caggcgatcc tgccctggtg gcaaccgtgc accctagctc cctgctgaga
     8221 ggcccaaagg aggagcggga atccgccttc gctgggctgg tggacgacct gagagtggct
     8281 gccgatgtga gacctaaaag aaccgccgac ggcagcgaat tcgagcccaa gaagaagagg
     8341 aaagtctaac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     8401 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     8461 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     8521 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     8581 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     8641 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     8701 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     8761 gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc
     8821 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     8881 aacgcgcggg aagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     8941 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     9001 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     9061 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     9121 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     9181 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     9241 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     9301 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     9361 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     9421 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     9481 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     9541 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     9601 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     9661 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgaca
     9721 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     9781 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     9841 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     9901 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     9961 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    10021 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    10081 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    10141 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    10201 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    10261 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    10321 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    10381 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    10441 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    10501 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    10561 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    10621 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    10681 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    10741 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    10801 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
    10861 cgggagatcg atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg
    10921 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
    10981 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
    11041 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
    11101 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
    11161 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
    11221 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
    11281 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
//
