LOCUS       sequence-148603-       11082 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Genome editing for gene pyrE (CAC-002) in Clostridium acetobutylicum
            ATCC 824.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11082)
  AUTHORS   Li Q, Chen J, Minton NP, Zhang Y, Wen Z, Liu J, Yang H, Zeng Z, Ren 
            X, Yang J, Gu Y, Jiang W, Jiang Y, Yang S
  TITLE     CRISPR-based genome editing and expression control systems in 
            Clostridium acetobutylicum and Clostridium beijerinckii.
  JOURNAL   Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053.
  PUBMED    27213844
REFERENCE   2  (bases 1 to 11082)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            J. 2016 May 23. doi: 10.1002/biot.201600053."
COMMENT     Sequence Label: pNICKclos1.0
FEATURES             Location/Qualifiers
     source          1..11082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..48
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        69..144
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(2304..2330,2331..2333,2334..6410)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6422..6439)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6422..6438)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6431..6453)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6647..6666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6766..6788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6826..6844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6912..7016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7017..7085,7086..7877)
                     /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     complement(7235..7254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8048..8636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8537..8556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8790..8807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8924..8945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8960..8965,8966..8983,8984..8990)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8998..9014
                     /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)."
     primer_bind     9003..9025
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9022..9038
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9022..9038
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agcttggctg cagttgacag ctagctcagt cctaggtata atactagtta tacttaaatc
       61 acaggctggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      121 gaaaaagtgg caccgagtcg gtgctttttt gcggccgcat tcttatgaaa attcaagtgc
      181 tctaactgta atttttccag gaataaatta tggtcatgaa aagcctcttc tgtattatgc
      241 aaggaaagca gctgcacaga ataatataga tattctatgc atcaactata agaataagat
      301 ttcttgggag gacataggaa aacaaagtat tgaagatgta gctattgaaa taaagggcat
      361 tataaaaaat gttattgaaa aaaagtatga taacatttat tttttaggga aaagtgttgg
      421 aacagaaata gctggatgta catcttctaa attaaatata gataatataa agtttcttta
      481 cttaacacct attgaagaga ctttaaagta tatagttgac agaaagtgca ttgtggtgtc
      541 aggcactaag gatgaatttt ttaaaagtga atgtataaat aaaatcagag aaaataaaaa
      601 tgttaaattg atgttgtttg agggagctaa tcattctctt gagattaagg atgatattat
      661 aagaagttta agaattttag ataacgtagc acaattgtat ttggacttct ttaaataaaa
      721 acatggaggg ctaaaaatgg aacaatacaa acaagaattt atagaattta tggtggagag
      781 taatgtactt acctttgggg atttcataac taaaagcggc agaagaacac cattttttat
      841 aaatacaggt aactacaaga caggtaatca attaaataag ttggctaagt tttatgctaa
      901 agcaatatat gataattttg gagatgattt tgatatttta tttgggcctg catataaagg
      961 aataccttta agtgtttcag tagctatggc acttgataat atttatggaa ttaatgcagc
     1021 ttattgttca aatagaaaag aagttaaaga tcacggtgat aagggaatac ttcttggagc
     1081 aaagcttgaa gaaggagaca gagttataat tgtagaagat gtcacaacag ctggtacatc
     1141 agtatacgaa acaatgccga tatcaggttg atgtaaaggg aatcataata tcagtggata
     1201 gaatggaaag aggtaaggga gataagagtg ccttaactga acttaaagaa aagtttggat
     1261 ttaaaacatg ttctattgtt actatggaag aggtagtaga atatttgtat aagaaaaata
     1321 tcaatggcaa agtaatcata gatgataaaa tgaaagatag aattaatgag tactataaag
     1381 agtatggagt aaaatagtaa aataaatgtg cctcaactta gatgttaagg cacatttatt
     1441 ttatatatta ttcatgtttt gaaacatttt tatcttttgt gtattttacg tgtagtaatt
     1501 tgtgagcaag tccttcacct ggttttccaa agtagctatc atacatttta ataatagctg
     1561 gattatcatg tgactttctc tttgaaagaa catttttatc ttggttgtat aatactgatg
     1621 ctcttagttt tctgtaatca acattttctc tatcaagagc atttacgtga ggttgacctc
     1681 caccatttat acatccacca gggcaagcca ttacttctat aaagtgatat tgtttttcgt
     1741 tcatttttcc agatttcata aactcgaaga agttagaagc accatttata acagcaacgt
     1801 ttagtttatt tccagcaatt tcaacttccg cttcttttat gcctttaaag cctcttactt
     1861 cagtgtaatc aacattttca agttctttat tttcagcaaa gtctttagct gatcttattg
     1921 cagcttccat aacgccaccg gttgcaccaa agatagctcc agcaccactg taagtaccca
     1981 tagcaggatc aacttcacca tcttcaagat ctgcaaattt aatttttgca tctttaatca
     2041 tttttgcaag ctctcttgta gttaaggatg catcaatatc tcttaagctg ttagtttcca
     2101 tgaaaggaat atctgcttca tattttttat cattacaagg catgatagta actgtataaa
     2161 catcctcgag ctgcagtata aaatataaat aattttctaa aaaacttaac ttcatgtgaa
     2221 aagtttgtta aaatataaat gagcacgtta atcatttaac atagataatt aaatagtaaa
     2281 agggagtgtc gaggatcccc gggatggata agaaatactc aataggctta gctatcggca
     2341 caaatagcgt cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca
     2401 aggttctggg aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat
     2461 ttgacagtgg agagacagcg gaagcgactc gtctcaaacg gacagctcgt agaaggtata
     2521 cacgtcggaa gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag
     2581 tagatgatag tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc
     2641 atgaacgtca tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaatatc
     2701 caactatcta tcatctgcga aaaaaattgg tagattctac tgataaagcg gatttgcgct
     2761 taatctattt ggccttagcg catatgatta agtttcgtgg tcattttttg attgagggag
     2821 atttaaatcc tgataatagt gatgtggaca aactatttat ccagttggta caaacctaca
     2881 atcaattatt tgaagaaaac cctattaacg caagtggagt agatgctaaa gcgattcttt
     2941 ctgcacgatt gagtaaatca agacgattag aaaatctcat tgctcagctc cccggtgaga
     3001 agaaaaatgg cttatttggg aatctcattg ctttgtcatt gggtttgacc cctaatttta
     3061 aatcaaattt tgatttggca gaagatgcta aattacagct ttcaaaagat acttacgatg
     3121 atgatttaga taatttattg gcgcaaattg gagatcaata tgctgatttg tttttggcag
     3181 ctaagaattt atcagatgct attttacttt cagatatcct aagagtaaat actgaaataa
     3241 ctaaggctcc cctatcagct tcaatgatta aacgctacga tgaacatcat caagacttga
     3301 ctcttttaaa agctttagtt cgacaacaac ttccagaaaa gtataaagaa atcttttttg
     3361 atcaatcaaa aaacggatat gcaggttata ttgatggggg agctagccaa gaagaatttt
     3421 ataaatttat caaaccaatt ttagaaaaaa tggatggtac tgaggaatta ttggtgaaac
     3481 taaatcgtga agatttgctg cgcaagcaac ggacctttga caacggctct attccccatc
     3541 aaattcactt gggtgagctg catgctattt tgagaagaca agaagacttt tatccatttt
     3601 taaaagacaa tcgtgagaag attgaaaaaa tcttgacttt tcgaattcct tattatgttg
     3661 gtccattggc gcgtggcaat agtcgttttg catggatgac tcggaagtct gaagaaacaa
     3721 ttaccccatg gaattttgaa gaagttgtcg ataaaggtgc ttcagctcaa tcatttattg
     3781 aacgcatgac aaactttgat aaaaatcttc caaatgaaaa agtactacca aaacatagtt
     3841 tgctttatga gtattttacg gtttataacg aattgacaaa ggtcaaatat gttactgaag
     3901 gaatgcgaaa accagcattt ctttcaggtg aacagaagaa agccattgtt gatttactct
     3961 tcaaaacaaa tcgaaaagta accgttaagc aattaaaaga agattatttc aaaaaaatag
     4021 aatgttttga tagtgttgaa atttcaggag ttgaagatag atttaatgct tcattaggta
     4081 cctaccatga tttgctaaaa attattaaag ataaagattt tttggataat gaagaaaatg
     4141 aagatatctt agaggatatt gttttaacat tgaccttatt tgaagatagg gagatgattg
     4201 aggaaagact taaaacatat gctcacctct ttgatgataa ggtgatgaaa cagcttaaac
     4261 gtcgccgtta tactggttgg ggacgtttgt ctcgaaaatt gattaatggt attagggata
     4321 agcaatctgg caaaacaata ttagattttt tgaaatcaga tggttttgcc aatcgcaatt
     4381 ttatgcagct gatccatgat gatagtttga catttaaaga agacattcaa aaagcacaag
     4441 tgtctggaca aggcgatagt ttacatgaac atattgcaaa tttagctggt agccctgcta
     4501 ttaaaaaagg tattttacag actgtaaaag ttgttgatga attggtcaaa gtaatggggc
     4561 ggcataagcc agaaaatatc gttattgaaa tggcacgtga aaatcagaca actcaaaagg
     4621 gccagaaaaa ttcgcgagag cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa
     4681 gtcagattct taaagagcat cctgttgaaa atactcaatt gcaaaatgaa aagctctatc
     4741 tctattatct ccaaaatgga agagacatgt atgtggacca agaattagat attaatcgtt
     4801 taagtgatta tgatgtcgat cacattgttc cacaaagttt ccttaaagac gattcaatag
     4861 acaataaggt cttaacgcgt tctgataaaa atcgtggtaa atcggataac gttccaagtg
     4921 aagaagtagt caaaaagatg aaaaactatt ggagacaact tctaaacgcc aagttaatca
     4981 ctcaacgtaa gtttgataat ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata
     5041 aagctggttt tatcaaacgc caattggttg aaactcgcca aatcactaag catgtggcac
     5101 aaattttgga tagtcgcatg aatactaaat acgatgaaaa tgataaactt attcgagagg
     5161 ttaaagtgat taccttaaaa tctaaattag tttctgactt ccgaaaagat ttccaattct
     5221 ataaagtacg tgagattaac aattaccatc atgcccatga tgcgtatcta aatgccgtcg
     5281 ttggaactgc tttgattaag aaatatccaa aacttgaatc ggagtttgtc tatggtgatt
     5341 ataaagttta tgatgttcgt aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa
     5401 ccgcaaaata tttcttttac tctaatatca tgaacttctt caaaacagaa attacacttg
     5461 caaatggaga gattcgcaaa cgccctctaa tcgaaactaa tggggaaact ggagaaattg
     5521 tctgggataa agggcgagat tttgccacag tgcgcaaagt attgtccatg ccccaagtca
     5581 atattgtcaa gaaaacagaa gtacagacag gcggattctc caaggagtca attttaccaa
     5641 aaagaaattc ggacaagctt attgctcgta aaaaagactg ggatccaaaa aaatatggtg
     5701 gttttgatag tccaacggta gcttattcag tcctagtggt tgctaaggtg gaaaaaggga
     5761 aatcgaagaa gttaaaatcc gttaaagagt tactagggat cacaattatg gaaagaagtt
     5821 cctttgaaaa aaatccgatt gactttttag aagctaaagg atataaggaa gttaaaaaag
     5881 acttaatcat taaactacct aaatatagtc tttttgagtt agaaaacggt cgtaaacgga
     5941 tgctggctag tgccggagaa ttacaaaaag gaaatgagct ggctctgcca agcaaatatg
     6001 tgaatttttt atatttagct agtcattatg aaaagttgaa gggtagtcca gaagataacg
     6061 aacaaaaaca attgtttgtg gagcagcata agcattattt agatgagatt attgagcaaa
     6121 tcagtgaatt ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg
     6181 catataacaa acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat
     6241 ttacgttgac gaatcttgga gctcccgctg cttttaaata ttttgataca acaattgatc
     6301 gtaaacgata tacgtctaca aaagaagttt tagatgccac tcttatccat caatccatca
     6361 ctggtcttta tgaaacacgc attgatttga gtcagctagg aggtgactga cccgggaatt
     6421 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     6481 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     6541 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     6601 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     6661 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8161 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     8221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8701 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8761 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8821 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8881 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8941 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     9001 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctttggc
     9061 taacacacac gccattccaa ccaatagttt tctcggcata aagccatgct ctgacgctta
     9121 aatgcactaa tgccttaaaa aaacattaaa gtctaacaca ctagacttat ttacttcgta
     9181 attaagtcgt taaaccgtgt gctctacgac caaaagtata aaacctttaa gaactttctt
     9241 ttttcttgta aaaaaagaaa ctagataaat ctctcatatc ttttattcaa taatcgcatc
     9301 agattgcagt ataaatttaa cgatcactca tcatgttcat atttatcaga gctcgtgcta
     9361 taattatact aattttataa ggaggaaaaa ataaagaggg ttataatgaa cgagaaaaat
     9421 ataaaacaca gtcaaaactt tattacttca aaacataata tagataaaat aatgacaaat
     9481 ataagattaa atgaacatga taatatcttt gaaatcggct caggaaaagg gcattttacc
     9541 cttgaattag tacagaggtg taatttcgta actgccattg aaatagacca taaattatgc
     9601 aaaactacag aaaataaact tgttgatcac gataatttcc aagttttaaa caaggatata
     9661 ttgcagttta aatttcctaa aaaccaatcc tataaaatat ttggtaatat accttataac
     9721 ataagtacgg atataatacg caaaattgtt tttgatagta tagctgatga gatttattta
     9781 atcgtggaat acgggtttgc taaaagatta ttaaatacaa aacgctcatt ggcattattt
     9841 ttaatggcag aagttgatat ttctatatta agtatggttc caagagaata ttttcatcct
     9901 aaacctaaag tgaatagctc acttatcaga ttaaatagaa aaaaatcaag aatatcacac
     9961 aaagataaac agaagtataa ttatttcgtt atgaaatggg ttaacaaaga atacaagaaa
    10021 atatttacaa aaaatcaatt taacaattcc ttaaaacatg caggaattga cgatttaaac
    10081 aatattagct ttgaacaatt cttatctctt ttcaatagct ataaattatt taataagtaa
    10141 gttaagggat gcataaactg catcccttaa cttgtttttc gtgtacctat tttttgtgaa
    10201 tcgattatgt cttttgcgca ttcacttctt ttctatataa atatgagcga agcgaataag
    10261 cgtcggaaaa gcagcaaaaa gtttcctttt tgctgttgga gcatgggggt tcagggggtg
    10321 cagtatctga cgtcaatgcc gagcgaaagc gagccgaagg gtagcattta cgttagataa
    10381 ccccctgata tgctccgacg ctttatatag aaaagaagat tcaactaggt aaaatcttaa
    10441 tataggttga gatgataagg tttataagga atttgtttgt tctaattttt cactcatttt
    10501 gttctaattt cttttaacaa atgttctttt ttttttagaa cagttatgat atagttagaa
    10561 tagtttaaaa taaggagtga gaaaaagatg aaagaaagat atggaacagt ctataaaggc
    10621 tctcagaggc tcatagacga agaaagtgga gaagtcatag aggtagacaa gttataccgt
    10681 aaacaaacgt ctggtaactt cgtaaaggca tatatagtgc aattaataag tatgttagat
    10741 atgattggcg gaaaaaaact taaaatcgtt aactatatcc tagataatgt ccacttaagt
    10801 aacaatacaa tgatagctac aacaagagaa atagcaaaag ctacaggaac aagtctacaa
    10861 acagtaataa caacacttaa aatcttagaa gaaggaaata ttataaaaag aaaaactgga
    10921 gtattaatgt taaaccctga actactaatg agaggcgacg accaaaaaca aaaatacctc
    10981 ttactcgaat ttgggaactt tgagcaagag gcaaatgaaa tagattgacc tcccaataac
    11041 accacgtagt tattgggagg tcaatctatg aaaatgcgat ta
//
