LOCUS       sequence-215976-       10601 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 10601)
  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 10601)
  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..10601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             676..696
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             703..723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             760..816
                     /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             898..954
                     /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             1036..1092
                     /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             1174..1230
                     /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             1312..1368
                     /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             1450..1506
                     /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             1588..1644
                     /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             1726..1782
                     /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             1864..1920
                     /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             2002..2058
                     /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..2118
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2125..2148,2149..2151,2152..6225)
                     /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             6256..6276
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6283..6300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6323..6340)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6329..6553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6599..7027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6686..6705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6896..6917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7036..7056)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7041..7370
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7221..7356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7283..7302
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7418..7465
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7484..7882
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8040..8161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8077..8096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8131..8150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8210..8226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8210..8226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8223..8245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8234..8250
                     /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(8258..8264,8265..8282,8283..8288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8303..8324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8441..8458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8612..9200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8692..8711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9371..10162,10163..10231))
                     /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     9994..10013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10232..10336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10411..10430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcacca tgggacccaa gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa
      721 gtggacggca ttggtagtgg gagcaacggc agcagcggag aggaactgct gagcaagaac
      781 taccacctgg aaaacgaggt ggccagactg aagaaaggct ctggctctgg cggaagcggt
      841 tctggcggat caggatctgg aagtggcggc tctggaagcg gaggttctgg atctggcgaa
      901 gaactgctgt ctaagaatta tcacctcgag aacgaagtgg ctcggctcaa gaaaggcagt
      961 ggcagcggag gaagtggatc cggcggtagt ggtagtggaa gcggcggatc cggctcaggc
     1021 ggatctggtt caggggaaga actcctctcc aaaaactacc atctcgagaa tgaggtcgcc
     1081 cgcctgaaaa aaggatcagg ttccggtggt tctggcagcg gtggttcagg ctcaggcagc
     1141 ggcggtagcg gtagcggtgg aagcggaagc ggagaagaac ttctcagcaa aaattaccac
     1201 ttggagaatg aagttgcaag actcaaaaaa ggttccggca gtggcggcag cggcagcggc
     1261 ggatctggta gtggatctgg tggcagtggt tcaggcggaa gtggtagcgg agaggaattg
     1321 ctctcaaaga actatcattt ggagaacgag gttgcacgct tgaagaaagg cagcggatca
     1381 ggcggatctg gcagcggtgg atctggttct ggatccggcg gctccggtag tggtggaagt
     1441 ggctctgggg aagaattgct tagcaagaat tatcatcttg aaaatgaggt tgccaggctt
     1501 aaaaaaggca gtggctccgg cggatccgga agcggaggca gcggatctgg atctggtggt
     1561 tcaggatctg gcggttctgg tagcggggaa gaactgttga gtaaaaacta tcaccttgag
     1621 aacgaggtcg caaggttgaa aaaaggatcc ggctctggcg gctccggaag tggcggatct
     1681 ggctccggta gcggaggatc aggatccggc ggaagcggat caggcgagga actgctttcc
     1741 aaaaattacc accttgaaaa cgaagtcgcc cgcctcaaga aaggttctgg cagcggaggc
     1801 tctggcagtg gtggtagcgg aagtggaagt ggtggcagtg gtagcggtgg atctggaagc
     1861 ggcgaggaac tcctgtcaaa gaattaccat ctcgaaaacg aggtcgcaag gctcaagaaa
     1921 ggctcaggat caggcggctc tggatccggc ggttctggtt ccggctcagg tggaagtgga
     1981 tctggcggct caggttccgg cgaagaattg ctttccaaga actaccattt ggaaaatgaa
     2041 gtcgctcgtt tgaagaaagg ttcaggctcc ggccagcggc cgcaaggtgg aggtggaccc
     2101 aagaagaagc gcaaggtgtc tagagacaag aaatactcta ttggactggc tatcgggaca
     2161 aactccgttg gctgggccgt cataaccgac gagtataagg tgccaagcaa gaaattcaag
     2221 gtgctgggta atactgaccg ccattcaatc aagaagaacc tgatcggagc actcctcttc
     2281 gactccggtg aaaccgctga agctactcgg ctgaagcgga ccgcaaggcg gagatacacc
     2341 cgccgcaaga atcggatatg ttatctgcaa gagatcttta gcaacgaaat ggctaaggtg
     2401 gacgactcct tctttcaccg cctggaagag agctttctgg tggaggagga taagaaacac
     2461 gagaggcacc ctatattcgg aaatatcgtg gatgaggtgg cttaccatga aaagtatcct
     2521 acaatctacc atctgaggaa gaagctggtg gacagcaccg ataaagcaga cctgaggctc
     2581 atctatctgg ccctggctca tatgataaag tttagaggac actttctgat cgagggcgac
     2641 ctgaatcccg ataattccga tgtggataaa ctcttcattc aactggtgca gacatataac
     2701 caactgttcg aggagaatcc cataaacgct tctggtgtgg atgccaaggc tattctgtcc
     2761 gctcggctgt ccaagtcacg cagactggag aatctgattg cccaactgcc aggagaaaag
     2821 aagaacggcc tgtttgggaa cctcatcgcc ctgagcctgg gcctgacacc taacttcaag
     2881 tccaattttg atctggccga agatgctaaa ctccagctct ccaaggacac ctatgacgat
     2941 gatctggaca acctgctcgc acagataggc gaccagtacg ccgatctctt tctggctgct
     3001 aagaatctct ccgacgccat tctgctgagc gacatactcc gggtcaacac tgagatcacc
     3061 aaagcacctc tgagcgcctc catgataaaa cgctatgatg aacaccatca agacctgact
     3121 ctgctcaaag ccctcgtgag gcaacagctg ccagagaagt acaaagagat attcttcgac
     3181 cagagcaaga atggatatgc cggatacatc gatggcggag catcacagga agaattttac
     3241 aagttcatca aaccaatcct cgagaagatg gacggtactg aagagctgct ggtgaagctg
     3301 aacagggagg acctgctgag gaagcagagg acctttgata atggctccat tccacatcag
     3361 atacacctgg gagagctgca tgcaatcctc cgcaggcagg aggatttcta tcctttcctg
     3421 aaggataacc gggagaagat agagaagatc ctgaccttca ggatccctta ttacgtcggc
     3481 cctctggcta gaggcaactc ccgcttcgct tggatgacca ggaaatctga ggagacaatt
     3541 actccttgga acttcgaaga ggtcgtggat aagggcgcaa gcgcccagtc attcatcgaa
     3601 cggatgacca atttcgataa gaacctgccc aacgagaagg tcctgcccaa acattcactc
     3661 ctgtacgagt atttcaccgt ctataacgag ctgactaaag tgaagtacgt gaccgagggc
     3721 atgaggaagc ctgccttcct gtccggagag cagaagaagg ctatcgttga tctgctcttc
     3781 aagactaata gaaaggtgac agtgaagcag ctcaaggagg attactttaa gaagatcgaa
     3841 tgctttgact cagtggaaat ctctggcgtg gaggaccgct ttaatgccag cctgggcact
     3901 taccatgatc tgctgaagat aatcaaagac aaagatttcc tcgataatga ggagaacgag
     3961 gacatcctgg aagatatcgt gctgaccctg actctgttcg aggatagaga gatgatcgaa
     4021 gagcgcctga agacctatgc ccatctgttt gacgataaag tcatgaaaca gctcaagcgg
     4081 cggcgctaca ctgggtgggg tagactctcc aggaaactca taaacggcat ccgcgacaaa
     4141 cagagcggaa agaccatcct ggatttcctg aaatccgacg gattcgctaa caggaacttc
     4201 atgcaactga ttcacgatga ctctctgaca tttaaagagg acatccagaa ggcacaggtg
     4261 agcggtcaag gcgacagcct gcacgagcac atcgccaacc tcgctggatc acccgccata
     4321 aagaagggaa tactgcagac agtcaaggtc gtggacgaac tcgtcaaagt gatgggtcgg
     4381 cacaagccag agaatatcgt tatcgaaatg gcaagggaga accaaaccac ccagaagggc
     4441 cagaagaact ctcgggaacg gatgaaaaga atcgaagagg gaattaagga gctgggatct
     4501 cagatactga aggagcaccc tgtggagaat acacagctcc agaacgagaa actctacctg
     4561 tactacctcc agaacgggcg ggacatgtac gttgaccagg aactcgacat caaccggctg
     4621 tccgattatg acgtggacca tattgttcca cagtccttcc tcaaagatga ctccattgac
     4681 aacaaggtgc tgaccagatc cgataagaat cgcggtaagt ctgacaatgt tccatcagaa
     4741 gaggtggtca agaagatgaa gaattactgg cggcagctcc tcaacgccaa actgatcacc
     4801 cagcggaagt ttgacaatct gactaaggca gaaagaggag gtctgagcga actcgacaag
     4861 gccggcttta ttaagaggca actggtcgaa acacgccaga ttaccaaaca cgtggcacaa
     4921 atcctcgact ctaggatgaa cactaagtac gatgagaacg ataagctgat cagggaagtg
     4981 aaagtgataa ctctgaagag caagctggtg tctgacttcc ggaaggactt tcaattctac
     5041 aaagttcgcg aaataaacaa ttaccatcat gctcacgatg cctatctcaa tgctgtcgtt
     5101 ggcaccgccc tgatcaagaa ataccctaaa ctggagtctg agttcgtgta cggtgactat
     5161 aaagtctacg atgtgaggaa gatgatagca aagtctgagc aagagattgg caaagccacc
     5221 gccaagtact tcttctactc taatatcatg aatttcttta agactgagat aaccctggct
     5281 aacggcgaaa tccggaagcg cccactgatc gaaacaaacg gagaaacagg agaaatcgtg
     5341 tgggataaag gcagggactt cgcaactgtg cggaaggtgc tgtccatgcc acaagtcaat
     5401 atcgtgaaga agaccgaagt gcagaccggc ggattctcaa aggagagcat cctgccaaag
     5461 cggaactctg acaagctgat cgccaggaag aaagattggg acccaaagaa gtatggcggt
     5521 ttcgattccc ctacagtggc ttattccgtt ctggtcgtgg caaaagtgga gaaaggcaag
     5581 tccaagaaac tcaagtctgt taaggagctg ctcggaatta ctattatgga gagatccagc
     5641 ttcgagaaga atccaatcga tttcctggaa gctaagggct ataaagaagt gaagaaagat
     5701 ctcatcatca aactgcccaa gtactctctc tttgagctgg agaatggtag gaagcggatg
     5761 ctggcctccg ccggagagct gcagaaagga aacgagctgg ctctgccctc caaatacgtg
     5821 aacttcctgt atctggcctc ccactacgag aaactcaaag gtagccctga agacaatgag
     5881 cagaagcaac tctttgttga gcaacataaa cactacctgg acgaaatcat tgaacagatt
     5941 agcgagttca gcaagcgggt tattctggcc gatgcaaacc tcgataaagt gctgagcgca
     6001 tataataagc acagggacaa gccaattcgc gaacaagcag agaatattat ccacctcttt
     6061 actctgacta atctgggcgc tcctgctgcc ttcaagtatt tcgatacaac tattgacagg
     6121 aagcggtaca cctctaccaa agaagttctc gatgccaccc tgatacacca gtcaattacc
     6181 ggactgtacg agactcgcat cgacctgtct cagctcggcg gcgacggttc tgaattcagc
     6241 ctgggcagcg gctcccccaa gaagaagcgc aaggtgaccg gtcatcatca ccatcaccat
     6301 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6541 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     6601 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6661 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6721 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6781 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6841 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6901 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6961 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7021 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     7081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     7141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7201 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     7261 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     7321 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     7381 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     7441 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc
     7501 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     7561 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     7621 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     7681 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     7741 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     7801 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     7861 tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga cacgtgctac
     7921 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7981 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     8041 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     8101 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     8161 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     8221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     8281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     8341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     8401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     8461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     8521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     8581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     8641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     8701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     8761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     8821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     8881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     8941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     9001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     9061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     9121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     9181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     9241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     9301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     9361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     9421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     9481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     9541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     9601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     9661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     9721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     9781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     9841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     9901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     9961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    10021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    10081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    10141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    10201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    10261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    10321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    10381 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
    10441 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    10501 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    10561 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
