LOCUS       sequence-386423-       12000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Base editing for making a pre-stop codon in mmoX using phenol 
            inducible system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12000)
  AUTHORS   Jeong J, Kim TH, Jang N, Ko M, Kim SK, Baek JI, Emelianov G, Rha E, 
            Kwon KK, Kim H, Lee EY, Lee DH, Lee H, Lee SG
  TITLE     A highly efficient and versatile genetic engineering toolkit for a 
            methanotroph-based biorefinery
  JOURNAL   Chem Eng J, 2023
REFERENCE   2  (bases 1 to 12000)
  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: "Chem Eng J,
            2023"
COMMENT     Sequence Label: pAWPSG-Po-nCas9-BE- Ptac-sgRNA(mmoX_1)
FEATURES             Location/Qualifiers
     source          1..12000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..648
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(912..1006)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     1017..1039
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             3151..3173
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3181..3867
                     /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(3916..3939,3940..3942,3943..8016)
                     /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             8029..8280
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        join(8321..8326,8327..8342,8343..8349)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_RNA        8362..8411
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_RNA        8455..8530
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      8573..8659
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(8812..9627)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(8812..9627)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     9535..9554
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9619..9638
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(9931..11079)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(11351..11722)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            11755..11864
                     /note="incP origin of transfer"
ORIGIN
        1 tccgtcgacc ctttccgacg ctcaccgggc tggttgccct cgccgctggg ctggcggccg
       61 tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga caccgcggcc
      121 gccggcgttg tggatacctc gcggaaaact tggccctcac tgacagatga ggggcggacg
      181 ttgacacttg aggggccgac tcacccggcg cggcgttgac agatgagggg caggctcgat
      241 ttcggccggc gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg
      301 cgagtttccc acagatgatg tggacaagcc tggggataag tgccctgcgg tattgacact
      361 tgaggggcgc gactactgac agatgagggg cgcgatcctt gacacttgag gggcagagtg
      421 ctgacagatg aggggcgcac ctattgacat ttgaggggct gtccacaggc agaaaatcca
      481 gcatttgcaa gggtttccgc ccgtttttcg gccaccgcta acctgtcttt taacctgctt
      541 ttaaaccaat atttataaac cttgttttta accagggctg cgccctgtgc gcgtgaccgc
      601 gcacgccgaa ggggggtgcc cccccttctc gaaccctccc ggcccgctaa cgcgggcctc
      661 ccatcccccc aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa aatggcagcc
      721 aagctgaccc gctagggacg tgaagtcgat tccttcgatg gttagcaatc aaagactcag
      781 agtgctgtct ggagcgtgaa tctaacggta cgtatctcga ttgctcggtc gctattcgca
      841 ctctgcgaaa gttcgtaccg ctcattcact aggttgcgaa aatttaaatc gtaattattg
      901 gggacccctg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
      961 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag actagtagcg
     1021 gataacaatt tcacacagga ttagccttcg atgccgattt tcttcagccg ataggccagt
     1081 gccggtcggc ttaggccgag caagcgcgcg gcaccggaga cgttttggtt ggcctgttgc
     1141 atggctttgc gcatcaggct ttcctcgatc tcgtcgaggc tcaggccgct gctcaacaac
     1201 tggctgatcc aactgccctg gccattgcct ggctgaatca gcacgccgtc cgacgacacc
     1261 cgctcgctgg cggtctgcgg ctcttccggc gcccgtggga acagcgcctg cacgctgatg
     1321 ctttcgttcg gatcggtgag gatgatgccg cgctcgatga cgttctccag ctcacggata
     1381 ttgcccggcc aactgtaatg caggcaggcc tccagggctt tgtctgaaag gccgagggtt
     1441 ctcttgccgt actcctggtg gaagcgctga aggaagtgct caaccagcag tggaatgtcc
     1501 tcgcggcgtt cgcgcaacgc cgggatcgcc accgggaaaa cgttcagccg gtagtacagg
     1561 tcggcgcgaa aacgcccggc cttgaccgct tcggccaggt cctcgtgggt ggctgcgata
     1621 acccttacgt cgatcttgcg cgtgcggttg tcgccaactc gctccagctc gccttcttgc
     1681 agcacgcgca gcagactggc ctgagcgcgc gggctgagtt cgatcacctc gtcaaggaag
     1741 atggtgccct tgtcggcccg ctcgaagcgg cccatgcgtg actgggtggc gccggtgaag
     1801 gcgccttttt ccacgccgaa caattcggac tcgatcaggt ccggcgggat cgccgcacag
     1861 ttcaccgcga caaagggctc ggcggcgcgt ttgctgcgca ggtgcacgct acgcgcgatg
     1921 acctccttgc cgaccccggt ctcgccaagc agcagcaccg agactttgcc ctgtgcggcc
     1981 ttgtccatca tattgcgcac ggtctggtag gccggggtct gaccgatgcc gtagtactgg
     2041 ccttcctgtt tgtcgaggtt ggtacgcagc gacaccagtt gcgattgcaa ctcgtagagt
     2101 tcctcgatga tggggtcgtt cttgaaatac tgtttgaagc tggcaacgtc gtcccactct
     2161 tcggccggct tgccaatgac ccggcacttg tcgccgccgc agccgcggca gctgacttcc
     2221 ttgaagatga tttcccggcc catgaacgcc gaggaatagg cgcaggcgta gccgagcaga
     2281 gtccagcaca ccgggtcttg catctgcccc aggtcggtct ggcagatttc cacctcgaag
     2341 gagtcgatcc actccatctc ggcatagaag cgcccgcatt ccttgtcgat atcgacctcg
     2401 gtggggcgga ccttgaccag acccttgagc gaatgcagct gcgggccggc gaggaacatg
     2461 tcgtactcgc tggcattcgg tctaagcttc ctggccagtt cggcatcctt caggccggac
     2521 tggtaaccct ggcgcaggaa gaagcccttg gcgcgttcga tgcccagggt attgaccatt
     2581 tcccggcgaa agctggccat tgctgaaacc tgcagcaaca gcatgcgttg ttcgccaagc
     2641 cagatcttgc cttccgtgct ctggaagtgg atcaggttgg tcaggtcctt gaaatcggag
     2701 tgctggattt caggcttgta cttgatcggc atgtaagcga ggcccctatt tatttttaga
     2761 tggggaaatc agggtcgccg ctatagcgca aggcaggcgg cgattccaga tggggtcatg
     2821 gggaaaatcg gcagtttttt cacctgcgcc agctccccat agccgacctc agtatctggc
     2881 aaaagtcaac caaatgatca atcgacggca gtgactttag tgctagagat cagccttcag
     2941 ctgcgccatg agcatttgct caagcggcct tgggcaattg atcaaatgct taaaaagcct
     3001 gcgcaagcgc ggcttaattt cgctcgctcc gatcattcta aaaattagaa acacattgaa
     3061 aaacaatacc ttgaagtcgg ttttcagacc ttggcacagc tgttgcactt tgtcctgcgc
     3121 aatccgccaa cctggagatc tagaaataat tttgtttaac tttaagaagg agatatacat
     3181 atgagctcag agactggccc agtggctgtg gaccccacat tgagacggcg gatcgagccc
     3241 catgagtttg aggtattctt cgatccgaga gagctccgca aggagacctg cctgctttac
     3301 gaaattaatt gggggggccg gcactccatt tggcgacata catcacagaa cactaacaag
     3361 cacgtcgaag tcaacttcat cgagaagttc acgacagaaa gatatttctg tccgaacaca
     3421 aggtgcagca ttacctggtt tctcagctgg agcccatgcg gcgaatgtag tagggccatc
     3481 actgaattcc tgtcaaggta tccccacgtc actctgttta tttacatcgc aaggctgtac
     3541 caccacgctg acccccgcaa tcgacaaggc ctgcgggatt tgatctcttc aggtgtgact
     3601 atccaaatta tgactgagca ggagtcagga tactgctgga gaaactttgt gaattatagc
     3661 ccgagtaatg aagcccactg gcctaggtat ccccatctgt gggtacgact gtacgttctt
     3721 gaactgtact gcatcatact gggcctgcct ccttgtctca acattctgag aaggaagcag
     3781 ccacagctga cattctttac catcgctctt cagtcttgtc attaccagcg actgccccca
     3841 cacattctct gggccaccgg gttgaaaagc ggcagcgaga ctcccgggac ctcagagtcc
     3901 gccacacccg aaagtgataa gaaatactca ataggcttag ctatcggcac aaatagcgtc
     3961 ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa ggttctggga
     4021 aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt tgacagtgga
     4081 gagacagcgg aagcgactcg tctcaaacgg acagctcgta gaaggtatac acgtcggaag
     4141 aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt agatgatagt
     4201 ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca tgaacgtcat
     4261 cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc aactatctat
     4321 catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt aatctatttg
     4381 gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga tttaaatcct
     4441 gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa tcaattattt
     4501 gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc tgcacgattg
     4561 agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc
     4621 ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa atcaaatttt
     4681 gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga tgatttagat
     4741 aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc taagaattta
     4801 tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac taaggctccc
     4861 ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac tcttttaaaa
     4921 gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga tcaatcaaaa
     4981 aacggatatg caggttatat tgatggggga gctagccaag aagaatttta taaatttatc
     5041 aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact aaatcgtgaa
     5101 gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca aattcacttg
     5161 ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt aaaagacaat
     5221 cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg tccattggcg
     5281 cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat taccccatgg
     5341 aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga acgcatgaca
     5401 aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt gctttatgag
     5461 tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa
     5521 ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt caaaacaaat
     5581 cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga atgttttgat
     5641 agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac ctaccatgat
     5701 ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga agatatctta
     5761 gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga ggaaagactt
     5821 aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg tcgccgttat
     5881 actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa gcaatctggc
     5941 aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt tatgcagctg
     6001 atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt gtctggacaa
     6061 ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat taaaaaaggt
     6121 attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg gcataagcca
     6181 gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat
     6241 tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag tcagattctt
     6301 aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct ctattatctc
     6361 caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt aagtgattat
     6421 gatgtcgatc acattgttcc acaaagtttc cttaaagacg attcaataga caataaggtc
     6481 ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga agaagtagtc
     6541 aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac tcaacgtaag
     6601 tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa agctggtttt
     6661 atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca aattttggat
     6721 agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt taaagtgatt
     6781 accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta taaagtacgt
     6841 gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt tggaactgct
     6901 ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta taaagtttat
     6961 gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac cgcaaaatat
     7021 ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc aaatggagag
     7081 attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt ctgggataaa
     7141 gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag
     7201 aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa aagaaattcg
     7261 gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg ttttgatagt
     7321 ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag
     7381 ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc ctttgaaaaa
     7441 aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga cttaatcatt
     7501 aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat gctggctagt
     7561 gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt gaatttttta
     7621 tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga acaaaaacaa
     7681 ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat cagtgaattt
     7741 tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc atataacaaa
     7801 catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt tacgttgacg
     7861 aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg taaacgatat
     7921 acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac tggtctttat
     7981 gaaacacgca ttgatttgag tcagctagga ggtgactctg gtggttctac taatctgtca
     8041 gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat cctcatgctc
     8101 ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact cgtgcacacc
     8161 gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc ccctgaatac
     8221 aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa gatgctctaa
     8281 ctgctgccac cgctgagcaa agctgctctg aaatgagctg ttgacaatta atcatcggct
     8341 cgtataatgt gtggactgca gctgtcaccg gatgtgcttt ccggtctgat gagtccgtga
     8401 ggacgaaaca gcctctacaa ataattttgt ttaaatgcac aggaagtgca ccgtgtttta
     8461 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     8521 gagtcggtgc tttttttgac ccggggcggc cgcctcgatt aattaaaggc atcaaataaa
     8581 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     8641 tctcctgagt aggacaaatc cgccgcccta gacagctggg cgcgccattg tcgggaagat
     8701 gcgtgatctg atccttccac tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg
     8761 tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa
     8821 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     8881 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     8941 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     9001 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     9061 tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     9121 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     9181 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     9241 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     9301 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     9361 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     9421 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     9481 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     9541 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca
     9601 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     9661 cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
     9721 agacacaacg tggctttcga cttccggcaa gctatacgcg ccctagaatt gtcaatttta
     9781 atcctctgtt tatcggcagt tcgtagagcg cgccgtgcgt cccgagcgat actgagcgaa
     9841 gcaagtgcgt cgagcagtgc ccgcttgttc ctgaaatgcc agtaaagcgc tggctgctga
     9901 acccccagcc ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt
     9961 gacccaggcg tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg
    10021 ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag
    10081 cttgagcggg tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg
    10141 cgacagcttg cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac
    10201 gacgatttcc tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac
    10261 gcggaagcgg tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc
    10321 catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt
    10381 gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat
    10441 aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg
    10501 cagctcgacg ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt
    10561 ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt
    10621 gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg
    10681 catttccttg atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac
    10741 ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt
    10801 gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc
    10861 cacggcggcc gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat
    10921 cttggccgta gcttgctgga ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg
    10981 catgacggtg cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag
    11041 ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc
    11101 cccgactcac gccggggcaa tgtgccctta ttcctgattt gacccgcctg gtgccttggt
    11161 gtccagataa tccaccttat cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt
    11221 ctcgtacttg gtattccgaa tcttgccctg cacgaatacc agctccgcga agtcgctctt
    11281 cttgatggag cgcatgggga cgtgcttggc aatcacgcgc accccccggc cgttttagcg
    11341 gctaaaaaag tcatggctct gccctcgggc ggaccacgcc catcatgacc ttgccaagct
    11401 cgtcctgctt ctcttcgatc ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg
    11461 ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc
    11521 cattgatgcg ggccagctcg cggacgtgct catagtccac gacgcccgtg attttgtagc
    11581 cctggccgac ggccagcagg taggcctaca ggctcatgcc ggccgccgcc gccttttcct
    11641 caatcgctct tcgttcgtct ggaaggcagt acaccttgat aggtgggctg cccttcctgg
    11701 ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg gtagccggcc
    11761 agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga
    11821 aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga
    11881 tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg
    11941 atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaaaga
//
