LOCUS       sequence-213093-       10635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses D10A-GCN4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10635)
  AUTHORS   Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang 
            L, Hou Y, Chen J, Chen J, Huang X
  TITLE     BE-PLUS: a new base editing tool with broadened editing window and 
            enhanced fidelity.
  JOURNAL   Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 
            10.1038/s41422-018-0052-4.
  PUBMED    29875396
REFERENCE   2  (bases 1 to 10635)
  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: "Cell Res. 
            2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 
            10.1038/s41422-018-0052-4."
COMMENT     Sequence Label: pST1374-D10A-GCN4
FEATURES             Location/Qualifiers
     source          1..10635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..930
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             940..963
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(970..993,994..996,997..5070)
                     /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             5101..5121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5128..5148
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5185..5241
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5323..5379
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5461..5517
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5599..5655
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5737..5793
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5875..5931
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6013..6069
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6151..6207
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6289..6345
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6427..6483
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6523..6543
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6550..6567
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6590..6607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6596..6820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6866..7294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6953..6972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7163..7184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7303..7323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7308..7638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7489..7624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7551..7570
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7686..7733
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7752..8150
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8308..8429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8345..8364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8399..8418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8478..8494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8478..8494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8491..8513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8502..8518
                     /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(8526..8532,8533..8550,8551..8556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8571..8592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8709..8726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8880..9468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8960..8979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9639..10430,10431..10499))
                     /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     10262..10281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10500..10604)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatgggac ctaagaaaaa gaggaaggtg gcggccgctg actacaagga tgacgacgat
      961 aaatctagag acaagaaata ctctattgga ctggctatcg ggacaaactc cgttggctgg
     1021 gccgtcataa ccgacgagta taaggtgcca agcaagaaat tcaaggtgct gggtaatact
     1081 gaccgccatt caatcaagaa gaacctgatc ggagcactcc tcttcgactc cggtgaaacc
     1141 gctgaagcta ctcggctgaa gcggaccgca aggcggagat acacccgccg caagaatcgg
     1201 atatgttatc tgcaagagat ctttagcaac gaaatggcta aggtggacga ctccttcttt
     1261 caccgcctgg aagagagctt tctggtggag gaggataaga aacacgagag gcaccctata
     1321 ttcggaaata tcgtggatga ggtggcttac catgaaaagt atcctacaat ctaccatctg
     1381 aggaagaagc tggtggacag caccgataaa gcagacctga ggctcatcta tctggccctg
     1441 gctcatatga taaagtttag aggacacttt ctgatcgagg gcgacctgaa tcccgataat
     1501 tccgatgtgg ataaactctt cattcaactg gtgcagacat ataaccaact gttcgaggag
     1561 aatcccataa acgcttctgg tgtggatgcc aaggctattc tgtccgctcg gctgtccaag
     1621 tcacgcagac tggagaatct gattgcccaa ctgccaggag aaaagaagaa cggcctgttt
     1681 gggaacctca tcgccctgag cctgggcctg acacctaact tcaagtccaa ttttgatctg
     1741 gccgaagatg ctaaactcca gctctccaag gacacctatg acgatgatct ggacaacctg
     1801 ctcgcacaga taggcgacca gtacgccgat ctctttctgg ctgctaagaa tctctccgac
     1861 gccattctgc tgagcgacat actccgggtc aacactgaga tcaccaaagc acctctgagc
     1921 gcctccatga taaaacgcta tgatgaacac catcaagacc tgactctgct caaagccctc
     1981 gtgaggcaac agctgccaga gaagtacaaa gagatattct tcgaccagag caagaatgga
     2041 tatgccggat acatcgatgg cggagcatca caggaagaat tttacaagtt catcaaacca
     2101 atcctcgaga agatggacgg tactgaagag ctgctggtga agctgaacag ggaggacctg
     2161 ctgaggaagc agaggacctt tgataatggc tccattccac atcagataca cctgggagag
     2221 ctgcatgcaa tcctccgcag gcaggaggat ttctatcctt tcctgaagga taaccgggag
     2281 aagatagaga agatcctgac cttcaggatc ccttattacg tcggccctct ggctagaggc
     2341 aactcccgct tcgcttggat gaccaggaaa tctgaggaga caattactcc ttggaacttc
     2401 gaagaggtcg tggataaggg cgcaagcgcc cagtcattca tcgaacggat gaccaatttc
     2461 gataagaacc tgcccaacga gaaggtcctg cccaaacatt cactcctgta cgagtatttc
     2521 accgtctata acgagctgac taaagtgaag tacgtgaccg agggcatgag gaagcctgcc
     2581 ttcctgtccg gagagcagaa gaaggctatc gttgatctgc tcttcaagac taatagaaag
     2641 gtgacagtga agcagctcaa ggaggattac tttaagaaga tcgaatgctt tgactcagtg
     2701 gaaatctctg gcgtggagga ccgctttaat gccagcctgg gcacttacca tgatctgctg
     2761 aagataatca aagacaaaga tttcctcgat aatgaggaga acgaggacat cctggaagat
     2821 atcgtgctga ccctgactct gttcgaggat agagagatga tcgaagagcg cctgaagacc
     2881 tatgcccatc tgtttgacga taaagtcatg aaacagctca agcggcggcg ctacactggg
     2941 tggggtagac tctccaggaa actcataaac ggcatccgcg acaaacagag cggaaagacc
     3001 atcctggatt tcctgaaatc cgacggattc gctaacagga acttcatgca actgattcac
     3061 gatgactctc tgacatttaa agaggacatc cagaaggcac aggtgagcgg tcaaggcgac
     3121 agcctgcacg agcacatcgc caacctcgct ggatcacccg ccataaagaa gggaatactg
     3181 cagacagtca aggtcgtgga cgaactcgtc aaagtgatgg gtcggcacaa gccagagaat
     3241 atcgttatcg aaatggcaag ggagaaccaa accacccaga agggccagaa gaactctcgg
     3301 gaacggatga aaagaatcga agagggaatt aaggagctgg gatctcagat actgaaggag
     3361 caccctgtgg agaatacaca gctccagaac gagaaactct acctgtacta cctccagaac
     3421 gggcgggaca tgtacgttga ccaggaactc gacatcaacc ggctgtccga ttatgacgtg
     3481 gaccatattg ttccacagtc cttcctcaaa gatgactcca ttgacaacaa ggtgctgacc
     3541 agatccgata agaatcgcgg taagtctgac aatgttccat cagaagaggt ggtcaagaag
     3601 atgaagaatt actggcggca gctcctcaac gccaaactga tcacccagcg gaagtttgac
     3661 aatctgacta aggcagaaag aggaggtctg agcgaactcg acaaggccgg ctttattaag
     3721 aggcaactgg tcgaaacacg ccagattacc aaacacgtgg cacaaatcct cgactctagg
     3781 atgaacacta agtacgatga gaacgataag ctgatcaggg aagtgaaagt gataactctg
     3841 aagagcaagc tggtgtctga cttccggaag gactttcaat tctacaaagt tcgcgaaata
     3901 aacaattacc atcatgctca cgatgcctat ctcaatgctg tcgttggcac cgccctgatc
     3961 aagaaatacc ctaaactgga gtctgagttc gtgtacggtg actataaagt ctacgatgtg
     4021 aggaagatga tagcaaagtc tgagcaagag attggcaaag ccaccgccaa gtacttcttc
     4081 tactctaata tcatgaattt ctttaagact gagataaccc tggctaacgg cgaaatccgg
     4141 aagcgcccac tgatcgaaac aaacggagaa acaggagaaa tcgtgtggga taaaggcagg
     4201 gacttcgcaa ctgtgcggaa ggtgctgtcc atgccacaag tcaatatcgt gaagaagacc
     4261 gaagtgcaga ccggcggatt ctcaaaggag agcatcctgc caaagcggaa ctctgacaag
     4321 ctgatcgcca ggaagaaaga ttgggaccca aagaagtatg gcggtttcga ttcccctaca
     4381 gtggcttatt ccgttctggt cgtggcaaaa gtggagaaag gcaagtccaa gaaactcaag
     4441 tctgttaagg agctgctcgg aattactatt atggagagat ccagcttcga gaagaatcca
     4501 atcgatttcc tggaagctaa gggctataaa gaagtgaaga aagatctcat catcaaactg
     4561 cccaagtact ctctctttga gctggagaat ggtaggaagc ggatgctggc ctccgccgga
     4621 gagctgcaga aaggaaacga gctggctctg ccctccaaat acgtgaactt cctgtatctg
     4681 gcctcccact acgagaaact caaaggtagc cctgaagaca atgagcagaa gcaactcttt
     4741 gttgagcaac ataaacacta cctggacgaa atcattgaac agattagcga gttcagcaag
     4801 cgggttattc tggccgatgc aaacctcgat aaagtgctga gcgcatataa taagcacagg
     4861 gacaagccaa ttcgcgaaca agcagagaat attatccacc tctttactct gactaatctg
     4921 ggcgctcctg ctgccttcaa gtatttcgat acaactattg acaggaagcg gtacacctct
     4981 accaaagaag ttctcgatgc caccctgata caccagtcaa ttaccggact gtacgagact
     5041 cgcatcgacc tgtctcagct cggcggcgac ggttctgaat tcagcctggg cagcggctcc
     5101 cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt
     5161 agtgggagca acggcagcag cggagaggaa ctgctgagca agaactacca cctggaaaac
     5221 gaggtggcca gactgaagaa aggctctggc tctggcggaa gcggttctgg cggatcagga
     5281 tctggaagtg gcggctctgg aagcggaggt tctggatctg gcgaagaact gctgtctaag
     5341 aattatcacc tcgagaacga agtggctcgg ctcaagaaag gcagtggcag cggaggaagt
     5401 ggatccggcg gtagtggtag tggaagcggc ggatccggct caggcggatc tggttcaggg
     5461 gaagaactcc tctccaaaaa ctaccatctc gagaatgagg tcgcccgcct gaaaaaagga
     5521 tcaggttccg gtggttctgg cagcggtggt tcaggctcag gcagcggcgg tagcggtagc
     5581 ggtggaagcg gaagcggaga agaacttctc agcaaaaatt accacttgga gaatgaagtt
     5641 gcaagactca aaaaaggttc cggcagtggc ggcagcggca gcggcggatc tggtagtgga
     5701 tctggtggca gtggttcagg cggaagtggt agcggagagg aattgctctc aaagaactat
     5761 catttggaga acgaggttgc acgcttgaag aaaggcagcg gatcaggcgg atctggcagc
     5821 ggtggatctg gttctggatc cggcggctcc ggtagtggtg gaagtggctc tggggaagaa
     5881 ttgcttagca agaattatca tcttgaaaat gaggttgcca ggcttaaaaa aggcagtggc
     5941 tccggcggat ccggaagcgg aggcagcgga tctggatctg gtggttcagg atctggcggt
     6001 tctggtagcg gggaagaact gttgagtaaa aactatcacc ttgagaacga ggtcgcaagg
     6061 ttgaaaaaag gatccggctc tggcggctcc ggaagtggcg gatctggctc cggtagcgga
     6121 ggatcaggat ccggcggaag cggatcaggc gaggaactgc tttccaaaaa ttaccacctt
     6181 gaaaacgaag tcgcccgcct caagaaaggt tctggcagcg gaggctctgg cagtggtggt
     6241 agcggaagtg gaagtggtgg cagtggtagc ggtggatctg gaagcggcga ggaactcctg
     6301 tcaaagaatt accatctcga aaacgaggtc gcaaggctca agaaaggctc aggatcaggc
     6361 ggctctggat ccggcggttc tggttccggc tcaggtggaa gtggatctgg cggctcaggt
     6421 tccggcgaag aattgctttc caagaactac catttggaaa atgaagtcgc tcgtttgaag
     6481 aaaggttcag gctccggcca gcggccgcaa ggtggaggtg gacccaagaa gaagcgcaag
     6541 gtgaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg
     6601 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6661 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6721 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6781 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     6841 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6901 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6961 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     7021 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     7081 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     7141 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     7201 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     7261 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     7321 ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat
     7381 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     7441 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     7501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     7561 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     7621 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     7681 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     7741 ggaactaaac catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa
     7801 cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct
     7861 ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg
     7921 cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg
     7981 tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgt cgacaggtgc
     8041 ttctcgatct gcatcctggg atcaaagcga tagtgaagga cagtgatgga cagccgacgg
     8101 cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg
     8161 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
     8221 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     8281 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     8341 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     8401 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     8461 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     8521 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     8581 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     8641 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     8701 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     8761 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     8821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     8881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     8941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     9001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     9061 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     9121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     9181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     9241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     9301 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     9361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     9421 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     9481 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     9541 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     9601 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     9661 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     9721 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     9781 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     9841 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     9901 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     9961 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
    10021 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
    10081 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
    10141 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
    10201 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
    10261 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
    10321 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
    10381 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
    10441 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
    10501 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
    10561 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
    10621 gtgccacctg acgtc
//
