LOCUS       sequence-472864-        9821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytosine base editor - spCas9n (D10A) fused to APOBEC1 and
            UGI, Include Golden Gate compatible cassette for sgRNA insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9821)
  AUTHORS   Lin H-C, Hsiao W-C, Hsu Y-C, Lin M-C, Hsu C-C, Zhang MM
  TITLE     Highly efficient CRISPR-Cas9 base editing in Bifidobacterium with 
            bypass of restriction modification systems.
  JOURNAL   Appl Environ Microbiol. 2025 Mar 10:e0198524. doi: 
            10.1128/aem.01985-24.
  PUBMED    40062897
REFERENCE   2  (bases 1 to 9821)
  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: "Appl 
            Environ Microbiol. 2025 Mar 10:e0198524. doi: 10.1128/aem.01985-24."
COMMENT     Sequence Label: cBEST4
FEATURES             Location/Qualifiers
     source          1..9821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..22
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        49..333
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
     CDS             334..1020
                     /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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(1069..1092,1093..1095,1096..5169)
                     /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             5182..5433
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     terminator      complement(5614..5662)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     protein_bind    5802..5820
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5976..5998
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5990..6007
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5991..6007
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6017..6036
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6017..6035
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6083..6101)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6122..6138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6122..6138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6135..6157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6146..6162
                     /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(6170..6176,6177..6194,6195..6200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6215..6236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        6278..6353
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      6375..6409
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(6451..7039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6531..6550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcaattaac cctcactaaa gggaacaaaa gctggagctc cagaattcgc tgctccttcg
       61 gtcggacgtg cgtctacggg caccttaccg cagccgtcgg ctgtgcgaca cggacggatc
      121 gggcgaactg gccgatgctg ggagaagcgc gctgctgtac ggcgcgcacc gggtgcggag
      181 cccctcggcg agcggtgtga aacttctgtg aatggcctgt tcggttgctt tttttatacg
      241 gctgccagat aaggcttgca gcatctgggc ggctaccgct atgatcgggg cgttcctgca
      301 attcttagtg cgagtatctg aaaggggata cgcatgtcct ccgagaccgg ccccgtggcc
      361 gtggacccga ccctgcgtcg ccgcatcgag ccgcacgagt tcgaggtgtt cttcgacccg
      421 cgcgagctgc gcaaggagac ctgcctgctg tacgagatca actggggcgg ccggcactcc
      481 atctggcgcc acacctcgca gaacaccaac aagcacgtcg aggtcaactt catcgagaag
      541 ttcaccaccg agcggtactt ctgcccgaac acccgctgct ccatcacctg gttcctgtcc
      601 tggtccccct gcggcgagtg ctcccgggcc atcaccgagt tcctgtcccg ctacccgcac
      661 gtcaccctgt tcatctacat cgcccggctg taccaccacg ccgacccgcg caaccggcag
      721 ggcctgcgcg acctgatctc ctccggcgtc accatccaga tcatgaccga gcaggagtcc
      781 ggctactgct ggcggaactt cgtcaactac tccccgtcca acgaggccca ctggccccgc
      841 tacccgcacc tgtgggtccg cctgtacgtc ctggagctgt actgcatcat cctgggcctg
      901 ccgccctgcc tgaacatcct gcgtcgcaag cagccgcagc tgaccttctt caccatcgcc
      961 ctccagtcct gccactacca gcgcctgccg ccccacatcc tgtgggccac cggcctgaag
     1021 tccggctccg agaccccggg cacctcggag tccgccaccc cggagtccga caagaagtac
     1081 tccatcggcc tcgcgatcgg caccaactcc gtgggctggg cggtcatcac cgacgagtac
     1141 aaggtcccct ccaagaagtt caaggtcctg ggcaacaccg accggcactc gatcaagaag
     1201 aacctgatcg gcgccctgct cttcgacagc ggcgagaccg ccgaggcgac ccgcctgaag
     1261 cggaccgcgc gtcgccgcta cacccggcgc aagaaccgca tctgctacct gcaggaaatc
     1321 ttctccaacg agatggccaa ggtggacgac tcgttcttcc accgcctgga ggagagcttc
     1381 ctggtggagg aggacaagaa gcacgagcgc cacccgatct tcggcaacat cgtggacgag
     1441 gtggcctacc acgagaagta ccccaccatc taccacctcc gcaagaagct ggtggactcg
     1501 accgacaagg cggacctgcg gctcatctac ctggccctcg cgcacatgat caagttccgc
     1561 ggccacttcc tcatcgaggg cgacctgaac ccggacaact ccgacgtgga caagctcttc
     1621 atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc
     1681 gtggacgcca aggcgatcct ctccgcgcgc ctgagcaagt cccggcgcct ggagaacctc
     1741 atcgcccagc tgccgggcga gaagaagaac ggcctcttcg gcaacctgat cgcgctgtcg
     1801 ctcggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc gaagctccag
     1861 ctgtccaagg acacctacga cgacgacctg gacaacctgc tcgcccagat cggcgaccag
     1921 tacgcggacc tcttcctggc cgcgaagaac ctctcggacg ccatcctgct cagcgacatc
     1981 ctgcgggtca acaccgagat caccaaggcc ccgctgtcgg cgagcatgat caagcggtac
     2041 gacgagcacc accaggacct gaccctgctc aaggccctcg tgcgccagca gctgcccgag
     2101 aagtacaagg aaatcttctt cgaccagtcc aagaacggct acgccggcta catcgacggc
     2161 ggcgcgtcgc aggaggagtt ctacaagttc atcaagccga tcctggagaa gatggacggc
     2221 accgaggagc tgctcgtcaa gctgaaccgc gaggacctgc tccgcaagca gcggaccttc
     2281 gacaacggct ccatcccgca ccagatccac ctgggcgagc tccacgccat cctccggcgc
     2341 caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc
     2401 ttccgcatcc cgtactacgt cggccccctg gcccgcggca actcccggtt cgcgtggatg
     2461 acccggaagt cggaggagac catcaccccg tggaacttcg aggaggtcgt ggacaagggc
     2521 gcgtccgcgc agtcgttcat cgagcgcatg accaacttcg acaagaacct cccgaacgag
     2581 aaggtcctgc ccaagcactc cctgctctac gagtacttca ccgtgtacaa cgagctgacc
     2641 aaggtcaagt acgtgaccga gggcatgcgg aagccggcct tcctgtcggg cgagcagaag
     2701 aaggcgatcg tggacctgct cttcaagacc aaccgcaagg tcaccgtgaa gcagctgaag
     2761 gaggactact tcaagaagat cgagtgcttc gactccgtcg agatcagcgg cgtggaggac
     2821 cgcttcaacg cctccctggg cacctaccac gacctgctca agatcatcaa ggacaaggac
     2881 ttcctcgaca acgaggagaa cgaggacatc ctggaggaca tcgtcctcac cctgaccctc
     2941 ttcgaggacc gcgagatgat cgaggagcgg ctcaagacct acgcccacct gttcgacgac
     3001 aaggtgatga agcagctgaa gcgtcgccgc tacaccggct ggggccgcct ctcccggaag
     3061 ctgatcaacg gcatccggga caagcagagc ggcaagacca tcctggactt cctcaagtcc
     3121 gacggcttcg ccaaccgcaa cttcatgcag ctcatccacg acgacagcct gaccttcaag
     3181 gaggacatcc agaaggccca ggtctcgggc cagggcgaca gcctccacga gcacatcgcc
     3241 aacctggcgg gctccccggc gatcaagaag ggcatcctcc agaccgtcaa ggtcgtggac
     3301 gagctggtca aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgg
     3361 gagaaccaga ccacccagaa gggccagaag aactcgcgcg agcggatgaa gcggatcgag
     3421 gagggcatca aggagctcgg cagccagatc ctgaaggagc acccggtcga gaacacccag
     3481 ctgcagaacg agaagctgta cctctactac ctgcagaacg gccgcgacat gtacgtggac
     3541 caggagctcg acatcaaccg gctgtccgac tacgacgtgg accacatcgt gccgcagtcc
     3601 ttcctgaagg acgactcgat cgacaacaag gtcctgaccc gctcggacaa gaaccggggc
     3661 aagtccgaca acgtgccctc ggaggaggtc gtgaagaaga tgaagaacta ctggcgccag
     3721 ctgctcaacg ccaagctcat cacccagcgc aagttcgaca acctgaccaa ggccgagcgg
     3781 ggcggcctga gcgagctcga caaggcgggc ttcatcaagc gccagctggt cgagacccgg
     3841 cagatcacca agcacgtggc ccagatcctg gactcccgga tgaacaccaa gtacgacgag
     3901 aacgacaagc tgatccgcga ggtcaaggtg atcaccctca agagcaagct ggtctccgac
     3961 ttccgcaagg acttccagtt ctacaaggtc cgggagatca acaactacca ccacgcccac
     4021 gacgcgtacc tgaacgccgt cgtgggcacc gcgctgatca agaagtaccc gaagctggag
     4081 tccgagttcg tctacggcga ctacaaggtc tacgacgtgc gcaagatgat cgccaagagc
     4141 gagcaggaga tcggcaaggc caccgcgaag tacttcttct actccaacat catgaacttc
     4201 ttcaagaccg agatcaccct ggccaacggc gagatccgca agcggcccct gatcgagacc
     4261 aacggcgaga ccggcgagat cgtctgggac aagggccgcg acttcgccac cgtccggaag
     4321 gtgctgtcga tgccgcaggt caacatcgtg aagaaaaccg aggtgcagac cggcggcttc
     4381 agcaaggagt ccatcctccc caagcgcaac agcgacaagc tgatcgcccg gaagaaggac
     4441 tgggacccga agaagtacgg cggcttcgac agccccaccg tcgcctactc cgtgctggtc
     4501 gtggcgaagg tcgagaaggg caagagcaag aagctgaagt ccgtgaagga gctgctcggc
     4561 atcaccatca tggagcgctc ctcgttcgag aagaacccga tcgacttcct ggaggccaag
     4621 ggctacaagg aggtcaagaa ggacctcatc atcaagctgc ccaagtacag cctgttcgag
     4681 ctggagaacg gccgcaagcg gatgctcgcc tccgcgggcg agctgcagaa gggcaacgag
     4741 ctggccctcc cgtcgaagta cgtcaacttc ctgtacctcg cgtcccacta cgagaagctg
     4801 aagggctcgc ccgaggacaa cgagcagaag cagctcttcg tggagcagca caagcactac
     4861 ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtcatcct ggccgacgcg
     4921 aacctcgaca aggtgctgtc cgcctacaac aagcaccgcg acaagccgat ccgggagcag
     4981 gcggagaaca tcatccacct gttcaccctc accaacctgg gcgcccccgc cgcgttcaag
     5041 tacttcgaca ccaccatcga ccgcaagcgg tacacctcca ccaaggaggt cctcgacgcg
     5101 accctgatcc accagagcat caccggcctg tacgagaccc gcatcgacct gtcccagctc
     5161 ggcggcgact ccggcggctc caccaacctg tccgacatca tcgagaagga gaccggcaag
     5221 cagctggtca tccaggagtc catcctgatg ctgccggagg aagtcgagga agtcatcggc
     5281 aacaagcccg agtccgacat cctggtccac accgcctacg acgagtccac cgacgagaac
     5341 gtcatgctgc tgacctcgga cgcccccgag tacaagccct gggccctggt catccaggac
     5401 tccaacggcg agaacaagat caagatgctg tgatctagag gatccccggg taccgagctc
     5461 ggattcccca gatctaaagt tttgtcgtct ttccagacgt tagtaaatga attttctgta
     5521 tgaggttttg ctaaacaact ttcaacagtt tcagcggagt gagaatagaa aggaacaact
     5581 aaaggaattg cgaataataa ttttttcacg ttgaaaatct ccaaaaaaaa aggctccaaa
     5641 aggagccttt aattgtatcg gtttatcagc ttgctttcga ggtgaatttc ttaaacagct
     5701 tgataccgat agttgcgccg acaatgacaa caaccatcgc ccacgcataa ccgatatatt
     5761 cgactagtct gttggctact ctatcattga tagtggtagg atccctatca gtgatagaga
     5821 taagtcttct cagccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     5881 ggaaggggga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5941 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6001 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
     6061 cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     6121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6241 cgttgcgctc actgcccgct tccaccggtg aagacatgtt ttagagctag aaatagcaag
     6301 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttac
     6361 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatc tagaggccag
     6421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg acccccccgt
     6721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6961 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7021 cagaaaaaaa ggatctcaag cggccgcatc ccattatgct ttggcagttt attcttgaca
     7081 tgtagtgagg gggctggtat aatcacatac ggccgataaa gcaagcatat aatattgcgt
     7141 ttcatcttta gaagcgaatt tcgccaatat tataattatc aaagagaggg gtggcaaacg
     7201 gtatttggca ttattaggtt aaaaaatgta gaaggagagt gaaacccatg aactttaata
     7261 aaattgattt agacaattgg aagagaaaag agatatttaa tcattatttg aaccaacaaa
     7321 cgacttttag tataaccaca gaaattgata ttagtgtttt ataccgaaac ataaaacaag
     7381 aaggatataa attttaccct gcatttattt tcttagtgac aagggtgata aactcaaata
     7441 cagcttttag aactggttac aatagcgacg gagagttagg ttattgggat aagttagagc
     7501 cactttatac aatttttgat ggtgtatcta aaacattctc tggtatttgg actcctgtaa
     7561 agaatgactt caaagagttt tatgatttat acctttctga tgtagagaaa tataatggtt
     7621 cggggaaatt gtttcccaaa acacctatac ctgaaaatgc tttttctctt tctattattc
     7681 catggacttc atttactggg tttaacttaa atatcaataa taatagtaat taccttctac
     7741 ccattattac agcaggaaaa ttcattaata aaggtaattc aatatattta ccgctatctt
     7801 tacaggtaca tcattctgtt tgtgatggtt atcatgcagg attgtttatg aactctattc
     7861 aggaattgtc agataggcct aatgactggc ttttataata tgagataatg ccgactgtac
     7921 tttttacagt cggttttcta atgtcactaa cctgccccgt tagttgatgc atgatcatca
     7981 taattctgtc tcattatcta gagtcgccat tggggaagaa gagcaggatt tgcctgcggt
     8041 tgaccgagac ggagtgggcg acgatgaacc ggatggcgtt cgcgtgcggt cagatggata
     8101 tctccatgga cacccacatg gatatctcca tggacaccca catggatatc tccatggaca
     8161 cccacatgga tatctccatc aaatcgaagg taagcaagct agacggttca tgtacatgaa
     8221 ccgaacgcag cacctccctc gccttgcggc gctggagcaa caacccgagt gccgaaggcc
     8281 gatggaagag gaaaaagccg caaggcgagg gaggtgctgc gttcgcgttt gggacaacgt
     8341 ttagcgtcag tcgaacagtg gaatctcgcc gggtagatgc ttcgatttct ccgccgcctc
     8401 gtcctcgcgc tccttgcgag ccctgtagca gcgttgacgc acggcaccct ggctcatgtt
     8461 gaaatgctcg gcggtctgcg gccatgtatg gccctcgtca tcgtgatatg agcggactgc
     8521 ttcgcgcata gcggcttgtt catcaatcca atcctcgcgc ttcattgcca cccagctctg
     8581 aatggtgcgc gtggagacgc cgtatgcttc agctaattca cgtgcagaca ccgggcgctt
     8641 cttcctcaaa gtcgtcctta ccattccagt gcctccttca tgacctgctg atatttgttc
     8701 tcaccgtgtt tgtgtcctcg tgcggattgg atggcgatga atgtggctgc gttggcaatg
     8761 gcaccgtcga gccacatgcg gctgcgggtg acgatccatt tgtggatgct cttggcgatg
     8821 tcctcgacct cacgcgcgtg cagcggatcg gggaacagcg agacgttgag ttcgtggcag
     8881 gtgcggcgca cgtataggcg caatgcctcg ctgctggcct ccgaacggtc gggcagccgc
     8941 cgcacgtagc ggtaggcgag cgtgcgggcc ttgtcgaaga gcgtgcagtt gcgtcccagc
     9001 ccctgcgtgc gggcgcgctt ggtgcgcttc cagctgacgg gcggcatgtc cccgtgttcc
     9061 tggagcgact gcacgagctg ttccaggtcg taggtgtccc actcgatgat gtccgacgcc
     9121 cacgccggat gctcggggtt cttcatcaga agtcccgaat agcccatgtc cccgtcgcag
     9181 gagcggcgca gtccctccgt ggtggcgtgc aggagcttca acggcttgag acgcgccaga
     9241 tcggttctgg gcaccggaaa ggtgagcacc cagcccgcgt gagcgtgccc gttggccggg
     9301 ttctccgcga tccagttcgg ccgcatgccc tcgtgctccc acagggccat cgccttggcg
     9361 ttcgcgtcgt cgatgtccac cacgatcgtg ttcaccagcg cggcggggtt cgcctcgatg
     9421 tgcttgcatt tgagcgcgtc ggcgcggctc ctgcggtgca gctgggagta gtcgtcgtcg
     9481 cagcacagcg ggcgttcggg cagccaccag ccttcgaacg tgcgcgacag ctcaagcgtc
     9541 ggctgcgaat actcatcgga catggctaca gcatacatcg tatttgtaca gtgaacaaat
     9601 acgcgggaac caagtcagat atttcagccc agcgaagcgt cggctgaacc cagcccagca
     9661 catcctgcac gaaaagccgc tggacggccc tcagccgctt ctccacgccg ctcacgtatg
     9721 cgcgagcctc tctgacctgt tcggccgcgt ggaggccact cacggacgtt tctgagccgt
     9781 ctgagccgtc ggcgtccgct gcgcgttccg caggctgagg c
//
