LOCUS       sequence-275477-       11185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for human codon optimized BE4max 
            C-to-T base editor with 
            xCas9(3.7)(D10A/A262T/R324L/S409I/E480K/E543D/M694I/E1219V) and 
            P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11185)
  AUTHORS   Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
  TITLE     Unconstrained genome targeting with near-PAMless engineered 
            CRISPR-Cas9 variants.
  JOURNAL   Science. 2020 Apr 17;368(6488):290-296. doi: 
            10.1126/science.aba8853. Epub 2020 Mar 26.
  PUBMED    32217751
REFERENCE   2  (bases 1 to 11185)
  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: 
            "10.1126/science.aba8853"; journalName: "Science"; date: 
            "2020-04-17- 17"; volume: "368"; issue: "6488"; pages: "290-296"
COMMENT     Sequence Label: pCAG-CBE4max-xCas9(3.7)-P2A-EGFP (RTW4637)
FEATURES             Location/Qualifiers
     source          1..11185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1805..1825
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1826..2509
                     /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             2606..6706
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 mutant with broad PAM
                     compatibility (Hu et al., 2018)"
                     /label=xCas9-3.7
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2606..6706
                     /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
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6737..6985
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7016..7264
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7307..7327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7346..7402
                     /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             join(7403..7405,7406..7408,7409..8122)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7403..7405,7406..7408,7409..8119)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7709..7728)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8056..8077
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(8209..8228)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8274..8329
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8328..8347)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8688..8704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8688..8704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8701..8723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8712..8728
                     /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(8736..8742,8743..8760,8761..8766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8781..8802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8861..9056
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8907..9042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9062..9196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9099..9118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9153..9172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9264..9281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9435..10023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9515..9534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10194..10985,10986..11054))
                     /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     10817..10836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11055..11159)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tcgccaccat gaaacggaca gccgacggaa gcgagttcga
     1801 gtcaccaaag aagaagcgga aagtctcctc agagactggg cctgtcgccg tcgatccaac
     1861 cctgcgccgc cggattgaac ctcacgagtt tgaagtgttc tttgaccccc gggagctgag
     1921 aaaggagaca tgcctgctgt acgagatcaa ctggggaggc aggcactcca tctggaggca
     1981 cacctctcag aacacaaata agcacgtgga ggtgaacttc atcgagaagt ttaccacaga
     2041 gcggtacttc tgccccaata ccagatgtag catcacatgg tttctgagct ggtccccttg
     2101 cggagagtgt agcagggcca tcaccgagtt cctgtccaga tatccacacg tgacactgtt
     2161 tatctacatc gccaggctgt atcaccacgc agacccaagg aataggcagg gcctgcgcga
     2221 tctgatcagc tccggcgtga ccatccagat catgacagag caggagtccg gctactgctg
     2281 gcggaacttc gtgaattatt ctcctagcaa cgaggcccac tggcctaggt acccacacct
     2341 gtgggtgcgc ctgtacgtgc tggagctgta ttgcatcatc ctgggcctgc ccccttgtct
     2401 gaatatcctg cggagaaagc agccccagct gaccttcttt acaatcgccc tgcagtcttg
     2461 tcactatcag aggctgccac cccacatcct gtgggccaca ggcctgaagt ctggaggatc
     2521 tagcggagga tcctctggca gcgagacacc aggaacaagc gagtcagcaa caccagagag
     2581 cagtggcggc agcagcggcg gcagcgacaa gaagtacagc atcggcctgg ccatcggcac
     2641 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     2701 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     2761 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     2821 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     2881 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     2941 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     3001 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     3061 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     3121 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     3181 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     3241 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     3301 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     3361 gagcaacttc gacctggccg aggataccaa actgcagctg agcaaggaca cctacgacga
     3421 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     3481 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     3541 caaggccccc ctgagcgcct ctatgatcaa gctgtacgac gagcaccacc aggacctgac
     3601 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     3661 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     3721 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     3781 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcatca tcccccacca
     3841 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     3901 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     3961 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     4021 caccccctgg aacttcgaga aggtggtgga caagggcgct tccgcccaga gcttcatcga
     4081 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     4141 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     4201 aatgagaaag cccgccttcc tgagcggcga ccagaaaaag gccatcgtgg acctgctgtt
     4261 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     4321 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     4381 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     4441 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     4501 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     4561 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     4621 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     4681 catccagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     4741 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     4801 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     4861 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     4921 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     4981 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     5041 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     5101 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     5161 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     5221 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     5281 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     5341 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     5401 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     5461 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     5521 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     5581 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     5641 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     5701 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     5761 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     5821 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     5881 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     5941 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     6001 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     6061 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     6121 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     6181 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     6241 gctggcctct gccggcgtgc tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     6301 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     6361 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     6421 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     6481 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     6541 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     6601 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     6661 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgacagcg gcgggagcgg
     6721 cgggagcggg gggagcacta atctgagcga catcattgag aaggagactg ggaaacagct
     6781 ggtcattcag gagtccatcc tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa
     6841 gccagagtct gacatcctgg tgcacaccgc ctacgacgag tccacagatg agaatgtgat
     6901 gctgctgacc tctgacgccc ccgagtataa gccttgggcc ctggtcatcc aggattctaa
     6961 cggcgagaat aagatcaaga tgctgagcgg aggatccgga ggatctggag gcagcaccaa
     7021 cctgtctgac atcatcgaga aggagacagg caagcagctg gtcatccagg agagcatcct
     7081 gatgctgccc gaagaagtcg aagaagtgat cggaaacaag cctgagagcg atatcctggt
     7141 ccataccgcc tacgacgaga gtaccgacga aaatgtgatg ctgctgacat ccgacgcccc
     7201 agagtataag ccctgggctc tggtcatcca ggattccaac ggagagaaca aaatcaaaat
     7261 gctgtctggc ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag
     7321 gaaagtcgga ggcggtggaa gcggcgcaac aaacttctct ctgctgaaac aagccggaga
     7381 tgtcgaagag aatcctggac cgatggtgag caagggcgag gagctgttca ccggggtggt
     7441 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     7501 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     7561 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     7621 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     7681 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     7741 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     7801 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     7861 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     7921 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     7981 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     8041 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     8101 catggacgag ctgtacaagt gaagcggccg cactcctcag gtgcaggctg cctatcagaa
     8161 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     8221 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     8281 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     8341 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     8401 atatgccata tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga
     8461 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     8521 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     8581 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     8641 tcttatgaag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     8701 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8761 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8821 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     8881 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     8941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     9001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     9061 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     9121 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     9181 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     9241 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     9301 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     9361 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     9421 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     9481 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     9541 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     9601 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     9661 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     9721 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     9781 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     9841 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     9901 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     9961 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    10021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    10081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    10141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    10201 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    10261 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    10321 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    10381 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    10441 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    10501 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    10561 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    10621 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    10681 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    10741 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    10801 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    10861 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    10921 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    10981 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    11041 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    11101 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    11161 acatttcccc gaaaagtgcc acctg
//
