LOCUS       sequence-386428-       12319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express APOVEC1-nCas9(D10A)-UGI using phenol inducible system in 
            Methanotroph or E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12319)
  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 12319)
  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
FEATURES             Location/Qualifiers
     source          1..12319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      31..662
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(926..1020)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     RBS             3165..3187
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3195..3881
                     /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(3930..3953,3954..3956,3957..8030)
                     /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             8043..8294
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     terminator      8318..8389
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8405..8432
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             8500..8529
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             8545..8562
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8618..8635)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      8788..8834
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8906..8992
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(9145..9960)
                     /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(9145..9960)
                     /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     9868..9887
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9952..9971
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(10264..11412)
                     /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(11684..12055)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            12088..12197
                     /note="incP origin of transfer"
ORIGIN
        1 tgcagcggaa aagatccgtc gaccctttcc gacgctcacc gggctggttg ccctcgccgc
       61 tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc
      121 gagacaccgc ggccgccggc gttgtggata cctcgcggaa aacttggccc tcactgacag
      181 atgaggggcg gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt tgacagatga
      241 ggggcaggct cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa
      301 cgcctgattt tacgcgagtt tcccacagat gatgtggaca agcctgggga taagtgccct
      361 gcggtattga cacttgaggg gcgcgactac tgacagatga ggggcgcgat ccttgacact
      421 tgaggggcag agtgctgaca gatgaggggc gcacctattg acatttgagg ggctgtccac
      481 aggcagaaaa tccagcattt gcaagggttt ccgcccgttt ttcggccacc gctaacctgt
      541 cttttaacct gcttttaaac caatatttat aaaccttgtt tttaaccagg gctgcgccct
      601 gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc tcccggcccg
      661 ctaacgcggg cctcccatcc ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc
      721 caaaaatggc agccaagctg acccgctagg gacgtgaagt cgattccttc gatggttagc
      781 aatcaaagac tcagagtgct gtctggagcg tgaatctaac ggtacgtatc tcgattgctc
      841 ggtcgctatt cgcactctgc gaaagttcgt accgctcatt cactaggttg cgaaaattta
      901 aatcgtaatt attggggacc cctggattct caccaataaa aaacgcccgg cggcaaccga
      961 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1021 caagactagt agcggataac aatttcacac aggattagcc ttcgatgccg attttcttca
     1081 gccgataggc cagtgccggt cggcttaggc cgagcaagcg cgcggcaccg gagacgtttt
     1141 ggttggcctg ttgcatggct ttgcgcatca ggctttcctc gatctcgtcg aggctcaggc
     1201 cgctgctcaa caactggctg atccaactgc cctggccatt gcctggctga atcagcacgc
     1261 cgtccgacga cacccgctcg ctggcggtct gcggctcttc cggcgcccgt gggaacagcg
     1321 cctgcacgct gatgctttcg ttcggatcgg tgaggatgat gccgcgctcg atgacgttct
     1381 ccagctcacg gatattgccc ggccaactgt aatgcaggca ggcctccagg gctttgtctg
     1441 aaaggccgag ggttctcttg ccgtactcct ggtggaagcg ctgaaggaag tgctcaacca
     1501 gcagtggaat gtcctcgcgg cgttcgcgca acgccgggat cgccaccggg aaaacgttca
     1561 gccggtagta caggtcggcg cgaaaacgcc cggccttgac cgcttcggcc aggtcctcgt
     1621 gggtggctgc gataaccctt acgtcgatct tgcgcgtgcg gttgtcgcca actcgctcca
     1681 gctcgccttc ttgcagcacg cgcagcagac tggcctgagc gcgcgggctg agttcgatca
     1741 cctcgtcaag gaagatggtg cccttgtcgg cccgctcgaa gcggcccatg cgtgactggg
     1801 tggcgccggt gaaggcgcct ttttccacgc cgaacaattc ggactcgatc aggtccggcg
     1861 ggatcgccgc acagttcacc gcgacaaagg gctcggcggc gcgtttgctg cgcaggtgca
     1921 cgctacgcgc gatgacctcc ttgccgaccc cggtctcgcc aagcagcagc accgagactt
     1981 tgccctgtgc ggccttgtcc atcatattgc gcacggtctg gtaggccggg gtctgaccga
     2041 tgccgtagta ctggccttcc tgtttgtcga ggttggtacg cagcgacacc agttgcgatt
     2101 gcaactcgta gagttcctcg atgatggggt cgttcttgaa atactgtttg aagctggcaa
     2161 cgtcgtccca ctcttcggcc ggcttgccaa tgacccggca cttgtcgccg ccgcagccgc
     2221 ggcagctgac ttccttgaag atgatttccc ggcccatgaa cgccgaggaa taggcgcagg
     2281 cgtagccgag cagagtccag cacaccgggt cttgcatctg ccccaggtcg gtctggcaga
     2341 tttccacctc gaaggagtcg atccactcca tctcggcata gaagcgcccg cattccttgt
     2401 cgatatcgac ctcggtgggg cggaccttga ccagaccctt gagcgaatgc agctgcgggc
     2461 cggcgaggaa catgtcgtac tcgctggcat tcggtctaag cttcctggcc agttcggcat
     2521 ccttcaggcc ggactggtaa ccctggcgca ggaagaagcc cttggcgcgt tcgatgccca
     2581 gggtattgac catttcccgg cgaaagctgg ccattgctga aacctgcagc aacagcatgc
     2641 gttgttcgcc aagccagatc ttgccttccg tgctctggaa gtggatcagg ttggtcaggt
     2701 ccttgaaatc ggagtgctgg atttcaggct tgtacttgat cggcatgtaa gcgaggcccc
     2761 tatttatttt tagatgggga aatcagggtc gccgctatag cgcaaggcag gcggcgattc
     2821 cagatggggt catggggaaa atcggcagtt ttttcacctg cgccagctcc ccatagccga
     2881 cctcagtatc tggcaaaagt caaccaaatg atcaatcgac ggcagtgact ttagtgctag
     2941 agatcagcct tcagctgcgc catgagcatt tgctcaagcg gccttgggca attgatcaaa
     3001 tgcttaaaaa gcctgcgcaa gcgcggctta atttcgctcg ctccgatcat tctaaaaatt
     3061 agaaacacat tgaaaaacaa taccttgaag tcggttttca gaccttggca cagctgttgc
     3121 actttgtcct gcgcaatccg ccaacctgga gatctagaaa taattttgtt taactttaag
     3181 aaggagatat acatatgagc tcagagactg gcccagtggc tgtggacccc acattgagac
     3241 ggcggatcga gccccatgag tttgaggtat tcttcgatcc gagagagctc cgcaaggaga
     3301 cctgcctgct ttacgaaatt aattgggggg gccggcactc catttggcga catacatcac
     3361 agaacactaa caagcacgtc gaagtcaact tcatcgagaa gttcacgaca gaaagatatt
     3421 tctgtccgaa cacaaggtgc agcattacct ggtttctcag ctggagccca tgcggcgaat
     3481 gtagtagggc catcactgaa ttcctgtcaa ggtatcccca cgtcactctg tttatttaca
     3541 tcgcaaggct gtaccaccac gctgaccccc gcaatcgaca aggcctgcgg gatttgatct
     3601 cttcaggtgt gactatccaa attatgactg agcaggagtc aggatactgc tggagaaact
     3661 ttgtgaatta tagcccgagt aatgaagccc actggcctag gtatccccat ctgtgggtac
     3721 gactgtacgt tcttgaactg tactgcatca tactgggcct gcctccttgt ctcaacattc
     3781 tgagaaggaa gcagccacag ctgacattct ttaccatcgc tcttcagtct tgtcattacc
     3841 agcgactgcc cccacacatt ctctgggcca ccgggttgaa aagcggcagc gagactcccg
     3901 ggacctcaga gtccgccaca cccgaaagtg ataagaaata ctcaataggc ttagctatcg
     3961 gcacaaatag cgtcggatgg gcggtgatca ctgatgaata taaggttccg tctaaaaagt
     4021 tcaaggttct gggaaataca gaccgccaca gtatcaaaaa aaatcttata ggggctcttt
     4081 tatttgacag tggagagaca gcggaagcga ctcgtctcaa acggacagct cgtagaaggt
     4141 atacacgtcg gaagaatcgt atttgttatc tacaggagat tttttcaaat gagatggcga
     4201 aagtagatga tagtttcttt catcgacttg aagagtcttt tttggtggaa gaagacaaga
     4261 agcatgaacg tcatcctatt tttggaaata tagtagatga agttgcttat catgagaaat
     4321 atccaactat ctatcatctg cgaaaaaaat tggtagattc tactgataaa gcggatttgc
     4381 gcttaatcta tttggcctta gcgcatatga ttaagtttcg tggtcatttt ttgattgagg
     4441 gagatttaaa tcctgataat agtgatgtgg acaaactatt tatccagttg gtacaaacct
     4501 acaatcaatt atttgaagaa aaccctatta acgcaagtgg agtagatgct aaagcgattc
     4561 tttctgcacg attgagtaaa tcaagacgat tagaaaatct cattgctcag ctccccggtg
     4621 agaagaaaaa tggcttattt gggaatctca ttgctttgtc attgggtttg acccctaatt
     4681 ttaaatcaaa ttttgatttg gcagaagatg ctaaattaca gctttcaaaa gatacttacg
     4741 atgatgattt agataattta ttggcgcaaa ttggagatca atatgctgat ttgtttttgg
     4801 cagctaagaa tttatcagat gctattttac tttcagatat cctaagagta aatactgaaa
     4861 taactaaggc tcccctatca gcttcaatga ttaaacgcta cgatgaacat catcaagact
     4921 tgactctttt aaaagcttta gttcgacaac aacttccaga aaagtataaa gaaatctttt
     4981 ttgatcaatc aaaaaacgga tatgcaggtt atattgatgg gggagctagc caagaagaat
     5041 tttataaatt tatcaaacca attttagaaa aaatggatgg tactgaggaa ttattggtga
     5101 aactaaatcg tgaagatttg ctgcgcaagc aacggacctt tgacaacggc tctattcccc
     5161 atcaaattca cttgggtgag ctgcatgcta ttttgagaag acaagaagac ttttatccat
     5221 ttttaaaaga caatcgtgag aagattgaaa aaatcttgac ttttcgaatt ccttattatg
     5281 ttggtccatt ggcgcgtggc aatagtcgtt ttgcatggat gactcggaag tctgaagaaa
     5341 caattacccc atggaatttt gaagaagttg tcgataaagg tgcttcagct caatcattta
     5401 ttgaacgcat gacaaacttt gataaaaatc ttccaaatga aaaagtacta ccaaaacata
     5461 gtttgcttta tgagtatttt acggtttata acgaattgac aaaggtcaaa tatgttactg
     5521 aaggaatgcg aaaaccagca tttctttcag gtgaacagaa gaaagccatt gttgatttac
     5581 tcttcaaaac aaatcgaaaa gtaaccgtta agcaattaaa agaagattat ttcaaaaaaa
     5641 tagaatgttt tgatagtgtt gaaatttcag gagttgaaga tagatttaat gcttcattag
     5701 gtacctacca tgatttgcta aaaattatta aagataaaga ttttttggat aatgaagaaa
     5761 atgaagatat cttagaggat attgttttaa cattgacctt atttgaagat agggagatga
     5821 ttgaggaaag acttaaaaca tatgctcacc tctttgatga taaggtgatg aaacagctta
     5881 aacgtcgccg ttatactggt tggggacgtt tgtctcgaaa attgattaat ggtattaggg
     5941 ataagcaatc tggcaaaaca atattagatt ttttgaaatc agatggtttt gccaatcgca
     6001 attttatgca gctgatccat gatgatagtt tgacatttaa agaagacatt caaaaagcac
     6061 aagtgtctgg acaaggcgat agtttacatg aacatattgc aaatttagct ggtagccctg
     6121 ctattaaaaa aggtatttta cagactgtaa aagttgttga tgaattggtc aaagtaatgg
     6181 ggcggcataa gccagaaaat atcgttattg aaatggcacg tgaaaatcag acaactcaaa
     6241 agggccagaa aaattcgcga gagcgtatga aacgaatcga agaaggtatc aaagaattag
     6301 gaagtcagat tcttaaagag catcctgttg aaaatactca attgcaaaat gaaaagctct
     6361 atctctatta tctccaaaat ggaagagaca tgtatgtgga ccaagaatta gatattaatc
     6421 gtttaagtga ttatgatgtc gatcacattg ttccacaaag tttccttaaa gacgattcaa
     6481 tagacaataa ggtcttaacg cgttctgata aaaatcgtgg taaatcggat aacgttccaa
     6541 gtgaagaagt agtcaaaaag atgaaaaact attggagaca acttctaaac gccaagttaa
     6601 tcactcaacg taagtttgat aatttaacga aagctgaacg tggaggtttg agtgaacttg
     6661 ataaagctgg ttttatcaaa cgccaattgg ttgaaactcg ccaaatcact aagcatgtgg
     6721 cacaaatttt ggatagtcgc atgaatacta aatacgatga aaatgataaa cttattcgag
     6781 aggttaaagt gattacctta aaatctaaat tagtttctga cttccgaaaa gatttccaat
     6841 tctataaagt acgtgagatt aacaattacc atcatgccca tgatgcgtat ctaaatgccg
     6901 tcgttggaac tgctttgatt aagaaatatc caaaacttga atcggagttt gtctatggtg
     6961 attataaagt ttatgatgtt cgtaaaatga ttgctaagtc tgagcaagaa ataggcaaag
     7021 caaccgcaaa atatttcttt tactctaata tcatgaactt cttcaaaaca gaaattacac
     7081 ttgcaaatgg agagattcgc aaacgccctc taatcgaaac taatggggaa actggagaaa
     7141 ttgtctggga taaagggcga gattttgcca cagtgcgcaa agtattgtcc atgccccaag
     7201 tcaatattgt caagaaaaca gaagtacaga caggcggatt ctccaaggag tcaattttac
     7261 caaaaagaaa ttcggacaag cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg
     7321 gtggttttga tagtccaacg gtagcttatt cagtcctagt ggttgctaag gtggaaaaag
     7381 ggaaatcgaa gaagttaaaa tccgttaaag agttactagg gatcacaatt atggaaagaa
     7441 gttcctttga aaaaaatccg attgactttt tagaagctaa aggatataag gaagttaaaa
     7501 aagacttaat cattaaacta cctaaatata gtctttttga gttagaaaac ggtcgtaaac
     7561 ggatgctggc tagtgccgga gaattacaaa aaggaaatga gctggctctg ccaagcaaat
     7621 atgtgaattt tttatattta gctagtcatt atgaaaagtt gaagggtagt ccagaagata
     7681 acgaacaaaa acaattgttt gtggagcagc ataagcatta tttagatgag attattgagc
     7741 aaatcagtga attttctaag cgtgttattt tagcagatgc caatttagat aaagttctta
     7801 gtgcatataa caaacataga gacaaaccaa tacgtgaaca agcagaaaat attattcatt
     7861 tatttacgtt gacgaatctt ggagctcccg ctgcttttaa atattttgat acaacaattg
     7921 atcgtaaacg atatacgtct acaaaagaag ttttagatgc cactcttatc catcaatcca
     7981 tcactggtct ttatgaaaca cgcattgatt tgagtcagct aggaggtgac tctggtggtt
     8041 ctactaatct gtcagatatt attgaaaagg agaccggtaa gcaactggtt atccaggaat
     8101 ccatcctcat gctcccagag gaggtggaag aagtcattgg gaacaagccg gaaagcgata
     8161 tactcgtgca caccgcctac gacgagagca ccgacgagaa tgtcatgctt ctgactagcg
     8221 acgcccctga atacaagcct tgggctctgg tcatacagga tagcaacggt gagaacaaga
     8281 ttaagatgct ctaactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc
     8341 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     8401 cactggctca ccttcgggtg ggcctttctg cgtttatacc tagggatata ttccgcttcc
     8461 tcggcgaccg gggcggccgc ctcgaggaag cttgggcccg aacaaaaact catctcagaa
     8521 gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc
     8581 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     8641 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     8701 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     8761 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     8821 gcctttcgtt ttatctgttg tttgtcggtg aactggcgcc caatacgcaa accgtgaaat
     8881 tgttatccgc tttaattaaa ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     8941 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc
     9001 ctagacagct gggcgcgcca ttgtcgggaa gatgcgtgat ctgatccttc cactcagcaa
     9061 aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     9121 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     9181 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     9241 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     9301 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     9361 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     9421 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     9481 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     9541 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     9601 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     9661 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     9721 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     9781 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     9841 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     9901 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
     9961 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
    10021 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt cgacttccgg
    10081 caagctatac gcgccctaga attgtcaatt ttaatcctct gtttatcggc agttcgtaga
    10141 gcgcgccgtg cgtcccgagc gatactgagc gaagcaagtg cgtcgagcag tgcccgcttg
    10201 ttcctgaaat gccagtaaag cgctggctgc tgaaccccca gccggaactg accccacaag
    10261 gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct
    10321 gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga
    10381 atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga
    10441 gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat
    10501 gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg
    10561 gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga
    10621 ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag
    10681 gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag
    10741 ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac
    10801 ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt
    10861 cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt
    10921 gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc
    10981 cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg
    11041 tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt
    11101 ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc
    11161 tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc
    11221 agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga
    11281 gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc
    11341 ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa
    11401 cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc
    11461 ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat
    11521 gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc
    11581 ctgcacgaat accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt
    11641 ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg
    11701 ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca
    11761 gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga
    11821 gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt
    11881 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggcct
    11941 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc
    12001 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
    12061 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
    12121 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
    12181 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
    12241 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
    12301 tgaccccgaa gcagggtta
//
