LOCUS       sequence-213108-       10602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GCN4-D10A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10602)
  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 10602)
  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-GCN4-D10A
FEATURES             Location/Qualifiers
     source          1..10602
                     /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             937..957
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             994..1050
                     /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             1132..1188
                     /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             1270..1326
                     /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             1408..1464
                     /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             1546..1602
                     /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             1684..1740
                     /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             1822..1878
                     /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             1960..2016
                     /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             2098..2154
                     /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             2236..2292
                     /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             2332..2352
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2359..2382,2383..2385,2386..6459)
                     /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             6490..6510
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6517..6534
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6557..6574)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6563..6787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6833..7261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6920..6939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7130..7151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7270..7290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7275..7605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7456..7591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7518..7537
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7653..7700
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7719..8117
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8275..8396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8312..8331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8366..8385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8445..8461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8445..8461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8458..8480)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8469..8485
                     /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(8493..8499,8500..8517,8518..8523))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8538..8559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8676..8693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8847..9435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8927..8946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9606..10397,10398..10466))
                     /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     10229..10248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10467..10571)
                     /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 ccaagaaaaa acgcaaggtg gaagatccta agaaaaagcg gaaagtggac
      961 ggcattggta gtgggagcaa cggcagcagc ggagaggaac tgctgagcaa gaactaccac
     1021 ctggaaaacg aggtggccag actgaagaaa ggctctggct ctggcggaag cggttctggc
     1081 ggatcaggat ctggaagtgg cggctctgga agcggaggtt ctggatctgg cgaagaactg
     1141 ctgtctaaga attatcacct cgagaacgaa gtggctcggc tcaagaaagg cagtggcagc
     1201 ggaggaagtg gatccggcgg tagtggtagt ggaagcggcg gatccggctc aggcggatct
     1261 ggttcagggg aagaactcct ctccaaaaac taccatctcg agaatgaggt cgcccgcctg
     1321 aaaaaaggat caggttccgg tggttctggc agcggtggtt caggctcagg cagcggcggt
     1381 agcggtagcg gtggaagcgg aagcggagaa gaacttctca gcaaaaatta ccacttggag
     1441 aatgaagttg caagactcaa aaaaggttcc ggcagtggcg gcagcggcag cggcggatct
     1501 ggtagtggat ctggtggcag tggttcaggc ggaagtggta gcggagagga attgctctca
     1561 aagaactatc atttggagaa cgaggttgca cgcttgaaga aaggcagcgg atcaggcgga
     1621 tctggcagcg gtggatctgg ttctggatcc ggcggctccg gtagtggtgg aagtggctct
     1681 ggggaagaat tgcttagcaa gaattatcat cttgaaaatg aggttgccag gcttaaaaaa
     1741 ggcagtggct ccggcggatc cggaagcgga ggcagcggat ctggatctgg tggttcagga
     1801 tctggcggtt ctggtagcgg ggaagaactg ttgagtaaaa actatcacct tgagaacgag
     1861 gtcgcaaggt tgaaaaaagg atccggctct ggcggctccg gaagtggcgg atctggctcc
     1921 ggtagcggag gatcaggatc cggcggaagc ggatcaggcg aggaactgct ttccaaaaat
     1981 taccaccttg aaaacgaagt cgcccgcctc aagaaaggtt ctggcagcgg aggctctggc
     2041 agtggtggta gcggaagtgg aagtggtggc agtggtagcg gtggatctgg aagcggcgag
     2101 gaactcctgt caaagaatta ccatctcgaa aacgaggtcg caaggctcaa gaaaggctca
     2161 ggatcaggcg gctctggatc cggcggttct ggttccggct caggtggaag tggatctggc
     2221 ggctcaggtt ccggcgaaga attgctttcc aagaactacc atttggaaaa tgaagtcgct
     2281 cgtttgaaga aaggttcagg ctccggccag cggccgcaag gtggaggtgg acccaagaag
     2341 aagcgcaagg tgtctagaga caagaaatac tctattggac tggctatcgg gacaaactcc
     2401 gttggctggg ccgtcataac cgacgagtat aaggtgccaa gcaagaaatt caaggtgctg
     2461 ggtaatactg accgccattc aatcaagaag aacctgatcg gagcactcct cttcgactcc
     2521 ggtgaaaccg ctgaagctac tcggctgaag cggaccgcaa ggcggagata cacccgccgc
     2581 aagaatcgga tatgttatct gcaagagatc tttagcaacg aaatggctaa ggtggacgac
     2641 tccttctttc accgcctgga agagagcttt ctggtggagg aggataagaa acacgagagg
     2701 caccctatat tcggaaatat cgtggatgag gtggcttacc atgaaaagta tcctacaatc
     2761 taccatctga ggaagaagct ggtggacagc accgataaag cagacctgag gctcatctat
     2821 ctggccctgg ctcatatgat aaagtttaga ggacactttc tgatcgaggg cgacctgaat
     2881 cccgataatt ccgatgtgga taaactcttc attcaactgg tgcagacata taaccaactg
     2941 ttcgaggaga atcccataaa cgcttctggt gtggatgcca aggctattct gtccgctcgg
     3001 ctgtccaagt cacgcagact ggagaatctg attgcccaac tgccaggaga aaagaagaac
     3061 ggcctgtttg ggaacctcat cgccctgagc ctgggcctga cacctaactt caagtccaat
     3121 tttgatctgg ccgaagatgc taaactccag ctctccaagg acacctatga cgatgatctg
     3181 gacaacctgc tcgcacagat aggcgaccag tacgccgatc tctttctggc tgctaagaat
     3241 ctctccgacg ccattctgct gagcgacata ctccgggtca acactgagat caccaaagca
     3301 cctctgagcg cctccatgat aaaacgctat gatgaacacc atcaagacct gactctgctc
     3361 aaagccctcg tgaggcaaca gctgccagag aagtacaaag agatattctt cgaccagagc
     3421 aagaatggat atgccggata catcgatggc ggagcatcac aggaagaatt ttacaagttc
     3481 atcaaaccaa tcctcgagaa gatggacggt actgaagagc tgctggtgaa gctgaacagg
     3541 gaggacctgc tgaggaagca gaggaccttt gataatggct ccattccaca tcagatacac
     3601 ctgggagagc tgcatgcaat cctccgcagg caggaggatt tctatccttt cctgaaggat
     3661 aaccgggaga agatagagaa gatcctgacc ttcaggatcc cttattacgt cggccctctg
     3721 gctagaggca actcccgctt cgcttggatg accaggaaat ctgaggagac aattactcct
     3781 tggaacttcg aagaggtcgt ggataagggc gcaagcgccc agtcattcat cgaacggatg
     3841 accaatttcg ataagaacct gcccaacgag aaggtcctgc ccaaacattc actcctgtac
     3901 gagtatttca ccgtctataa cgagctgact aaagtgaagt acgtgaccga gggcatgagg
     3961 aagcctgcct tcctgtccgg agagcagaag aaggctatcg ttgatctgct cttcaagact
     4021 aatagaaagg tgacagtgaa gcagctcaag gaggattact ttaagaagat cgaatgcttt
     4081 gactcagtgg aaatctctgg cgtggaggac cgctttaatg ccagcctggg cacttaccat
     4141 gatctgctga agataatcaa agacaaagat ttcctcgata atgaggagaa cgaggacatc
     4201 ctggaagata tcgtgctgac cctgactctg ttcgaggata gagagatgat cgaagagcgc
     4261 ctgaagacct atgcccatct gtttgacgat aaagtcatga aacagctcaa gcggcggcgc
     4321 tacactgggt ggggtagact ctccaggaaa ctcataaacg gcatccgcga caaacagagc
     4381 ggaaagacca tcctggattt cctgaaatcc gacggattcg ctaacaggaa cttcatgcaa
     4441 ctgattcacg atgactctct gacatttaaa gaggacatcc agaaggcaca ggtgagcggt
     4501 caaggcgaca gcctgcacga gcacatcgcc aacctcgctg gatcacccgc cataaagaag
     4561 ggaatactgc agacagtcaa ggtcgtggac gaactcgtca aagtgatggg tcggcacaag
     4621 ccagagaata tcgttatcga aatggcaagg gagaaccaaa ccacccagaa gggccagaag
     4681 aactctcggg aacggatgaa aagaatcgaa gagggaatta aggagctggg atctcagata
     4741 ctgaaggagc accctgtgga gaatacacag ctccagaacg agaaactcta cctgtactac
     4801 ctccagaacg ggcgggacat gtacgttgac caggaactcg acatcaaccg gctgtccgat
     4861 tatgacgtgg accatattgt tccacagtcc ttcctcaaag atgactccat tgacaacaag
     4921 gtgctgacca gatccgataa gaatcgcggt aagtctgaca atgttccatc agaagaggtg
     4981 gtcaagaaga tgaagaatta ctggcggcag ctcctcaacg ccaaactgat cacccagcgg
     5041 aagtttgaca atctgactaa ggcagaaaga ggaggtctga gcgaactcga caaggccggc
     5101 tttattaaga ggcaactggt cgaaacacgc cagattacca aacacgtggc acaaatcctc
     5161 gactctagga tgaacactaa gtacgatgag aacgataagc tgatcaggga agtgaaagtg
     5221 ataactctga agagcaagct ggtgtctgac ttccggaagg actttcaatt ctacaaagtt
     5281 cgcgaaataa acaattacca tcatgctcac gatgcctatc tcaatgctgt cgttggcacc
     5341 gccctgatca agaaataccc taaactggag tctgagttcg tgtacggtga ctataaagtc
     5401 tacgatgtga ggaagatgat agcaaagtct gagcaagaga ttggcaaagc caccgccaag
     5461 tacttcttct actctaatat catgaatttc tttaagactg agataaccct ggctaacggc
     5521 gaaatccgga agcgcccact gatcgaaaca aacggagaaa caggagaaat cgtgtgggat
     5581 aaaggcaggg acttcgcaac tgtgcggaag gtgctgtcca tgccacaagt caatatcgtg
     5641 aagaagaccg aagtgcagac cggcggattc tcaaaggaga gcatcctgcc aaagcggaac
     5701 tctgacaagc tgatcgccag gaagaaagat tgggacccaa agaagtatgg cggtttcgat
     5761 tcccctacag tggcttattc cgttctggtc gtggcaaaag tggagaaagg caagtccaag
     5821 aaactcaagt ctgttaagga gctgctcgga attactatta tggagagatc cagcttcgag
     5881 aagaatccaa tcgatttcct ggaagctaag ggctataaag aagtgaagaa agatctcatc
     5941 atcaaactgc ccaagtactc tctctttgag ctggagaatg gtaggaagcg gatgctggcc
     6001 tccgccggag agctgcagaa aggaaacgag ctggctctgc cctccaaata cgtgaacttc
     6061 ctgtatctgg cctcccacta cgagaaactc aaaggtagcc ctgaagacaa tgagcagaag
     6121 caactctttg ttgagcaaca taaacactac ctggacgaaa tcattgaaca gattagcgag
     6181 ttcagcaagc gggttattct ggccgatgca aacctcgata aagtgctgag cgcatataat
     6241 aagcacaggg acaagccaat tcgcgaacaa gcagagaata ttatccacct ctttactctg
     6301 actaatctgg gcgctcctgc tgccttcaag tatttcgata caactattga caggaagcgg
     6361 tacacctcta ccaaagaagt tctcgatgcc accctgatac accagtcaat taccggactg
     6421 tacgagactc gcatcgacct gtctcagctc ggcggcgacg gttctgaatt cagcctgggc
     6481 agcggctccc ccaagaagaa gcgcaaggtg accggtcatc atcaccatca ccattgagtt
     6541 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6601 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6661 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6721 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     6781 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     6841 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6901 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6961 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     7021 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     7081 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7141 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     7201 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7261 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc
     7321 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     7381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     7441 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     7501 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     7561 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     7621 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     7681 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag
     7741 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     7801 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     7861 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     7921 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     7981 gcccctgcgg acggtgtcga caggtgcttc tcgatctgca tcctgggatc aaagcgatag
     8041 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     8101 atgtgtggga gggctaagca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     8161 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     8221 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     8281 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     8341 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     8401 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     8461 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     8521 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     8581 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     8641 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     8701 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     8761 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     8821 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8881 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8941 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     9001 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     9061 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     9121 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     9181 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     9241 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     9301 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     9361 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     9421 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     9481 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     9541 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     9601 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     9661 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     9721 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     9781 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     9841 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     9901 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     9961 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    10021 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    10081 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    10141 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    10201 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    10261 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    10321 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    10381 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    10441 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    10501 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    10561 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
