LOCUS       sequence-403016-       10985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector for constitutive CRISPRi-mediated gene knockdown in S. 
            aureus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10985)
  AUTHORS   Long DR, Holmes EA, Lo HY, Penewit K, Almazan J, Hodgson T, Berger 
            NF, Bishop ZH, Lewis JD, Waalkes A, Wolter DJ, Salipante SJ
  TITLE     Clinical and in vitro models identify distinct adaptations enhancing
            Staphylococcus aureus pathogenesis in human macrophages.
  JOURNAL   PLoS Pathog. 2024 Jul 11;20(7):e1012394. doi: 
            10.1371/journal.ppat.1012394. eCollection 2024 Jul.
  PUBMED    38991026
REFERENCE   2  (bases 1 to 10985)
  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: "PLoS 
            Pathog."; date: "2024-07-11"; volume: "20(7):e1012394. doi"; pages: 
            " 10.1371/journal.ppat.1012394. eCollection 2024 Jul"
COMMENT     Sequence Label: pCRISPRi
FEATURES             Location/Qualifiers
     source          1..10985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..74
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(112..130)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        198..302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(303..371,372..1163)
                     /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(521..540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1334..1922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1823..1842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2140..2166,2167..2169,2170..4656,4657..4659,
                     4660..6246)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     EPILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     misc_RNA        complement(6736..6811)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(6825..7538,7539..7541))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
       61 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
      121 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      301 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      361 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      421 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      481 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      541 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      601 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      661 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      721 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      781 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      841 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      901 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      961 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggtt
     1021 cacgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1081 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1141 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1201 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1261 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1321 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1381 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1441 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1501 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1561 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1621 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1681 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1741 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1801 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1861 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1921 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1981 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2041 ctggcggccg ctgcatgcct gcagggctgg cggccgctgc atgatgacaa aaagagcaaa
     2101 attttgataa aatagtatta gtcgacaaag gaggcaaaaa tggataagaa atactcaata
     2161 ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
     2221 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
     2281 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca
     2341 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
     2401 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
     2461 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
     2521 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
     2581 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
     2641 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
     2701 ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat
     2761 gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct
     2821 cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt
     2881 ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca
     2941 aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct
     3001 gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga
     3061 gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa
     3121 catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat
     3181 aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct
     3241 agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag
     3301 gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac
     3361 ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa
     3421 gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga
     3481 attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg
     3541 aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca
     3601 gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta
     3661 ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc
     3721 aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc
     3781 attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat
     3841 tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt
     3901 aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg
     3961 gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa
     4021 gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg
     4081 atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt
     4141 aatggtatta gggataagca atctggcaaa acaatattag attttttgaa atcagatggt
     4201 tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac
     4261 attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta
     4321 gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg
     4381 gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat
     4441 cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt
     4501 atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa
     4561 aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa
     4621 ttagatatta atcgtttaag tgattatgat gtcgatgcca ttgttccaca aagtttcctt
     4681 aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg
     4741 gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta
     4801 aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt
     4861 ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc
     4921 actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat
     4981 aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga
     5041 aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg
     5101 tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag
     5161 tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa
     5221 gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa
     5281 acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg
     5341 gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg
     5401 tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag
     5461 gagccaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat
     5521 ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct
     5581 aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca
     5641 attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat
     5701 aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa
     5761 aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct
     5821 ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt
     5881 agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat
     5941 gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta
     6001 gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa
     6061 aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt
     6121 gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt
     6181 atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt
     6241 gactgaagta tattttagat gaagattatt tcttaataac taaaaatatg gtataatact
     6301 cttaataaat gcagtaatac aggggctttt caagactccc gggtaccgag ctcgaattca
     6361 ggcgcgccta ttctaaatgc ataataaata ctgataacat cttatatttt gtattatatt
     6421 ttgtattatc gttgacatgt ataattttga tatcaaaaac tgattttccc tctattattt
     6481 tcgagattta ttttcttaat tctctttaac aaactagaaa tattgtatat acaaaaaatt
     6541 ataaataata gatgaatagt ttaattatag gtgttcatca atcgaaaaag caacgtatct
     6601 tatttaaagt gcgttgcttt tttctcattt ataaggttaa ataattctca tatatcaagc
     6661 aaagtgacag gcgccaaata ataaaaaagc cggattaata atctggcttt ttatattctc
     6721 tggcagacta aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct
     6781 tattttaact tgctatttct agctctaaaa cgagaccatt atcattattt gtacaactcg
     6841 tccatgccgt gcgtgatgcc agctgctgtt acaaactcca ataaaaccat gtggtcacgt
     6901 ttctcgttcg ggtccttaga taatactgac tgtgtagata aatagtgatt atccggcaac
     6961 aacacaggtc cgtcaccgat tggcgtattc tgctggtagt ggtctgctaa ctgaacactt
     7021 ccatcttcta cgttatgtct tatcttaaaa tttgctttga taccattctt ctgtttgtca
     7081 gcagtgatgt acacattatg tgagttgaaa ttgtattcta acttatgtcc taagatattg
     7141 ccatcctctt tgaaatcaat accttttaac tcaattctat tcaccaaagt gtcgccctcg
     7201 aatttcactt cagctcttgt tttgtaagta ccgtcatctt tgaatgaaat tgtacgttct
     7261 tgtacataac cttccggcat agcacttttg aaaaagtcgt gtctcttcat gtggtctgga
     7321 taacgactga agcattggac tccgtatgtt aacgtcgtca ctaaagtagg ccatggaaca
     7381 ggcaacttac ctgtcgtaca aatgaacttt aatgtcaatt ttccgttagt cgcgtcgccc
     7441 tctccttcac ctcgcacaga aaatttatgg ccgttaacgt caccatccaa ttctactaag
     7501 atcggcacaa cacccgtgaa caactcttcg ccttttgaca taaataatca tcctcctaat
     7561 ttgatgcatg catcttgctc gatacatttg cccgataata tattgatatc taatctttat
     7621 ttattataga tatgttagtc ataattttgc attaaataag ttttattaaa tatatttaat
     7681 gctctattat ttagttaatt ataactaatt aaaaatgaga agtaaacaaa aaagtgtttc
     7741 ctgcaggggt ctcgagctta ttttaattaa tctctatcaa tgatagagtg ttcatatttt
     7801 ttttagtttt tcatgaaggc gcctttgcgg aaagagttaa taagttaaca gaagatgaac
     7861 caaaactaaa tggtttagca ggaaacttag ataaaaaaat gaatccagaa ttatattcag
     7921 aacaggaaca gcaacaagaa caacaaaaga atcaaaaacg agatagaggt atgcacttat
     7981 agaacatgca tttatgccga gaaaacttat tggttggaat gggctatgtg ttagctaact
     8041 tgttagcgag ttggttggac ttgaattggg attaatccca agaaagtacc aactcaacaa
     8101 cacataaagc cctgtaggtt ccgaccaata aggaaattgg aataaagcaa taaaaggagt
     8161 tgaagaaatg aaattcagag aagcctttga gaattttata acaagtaagt atgtacttgg
     8221 tgttttagta gttttaactg tttaccagat aatacaaatg cttaaataaa aaaagacttg
     8281 atctgattag accaaatctt ttgatagtgt tatattaata acaaaataaa aaggagtcgc
     8341 tcacgccctg accaaagttt gtgaacgaca tcattcaaag aaaaaaacac tgagttgttt
     8401 ttataatctt gtatatttag atattaaacg atatttaaat atacatcaag atatatattt
     8461 gggtgagcga ttccttaaac gaaattgaga ttaaggagtc gattttttat gtataaaaac
     8521 aatcatgcaa atcattcaaa tcatttggaa aatcacgatt tagacaattt ttctaaaacc
     8581 ggctactcta atagccggtt ggacgcacat actgtgtgca tatctgatcc aaaattaagt
     8641 tttgatgcaa tgacgatcgt tggaaatctc aaccgagaca acgctcaagc cctttctaaa
     8701 tttatgagtg tagagcccca aataagactt tgggatattc ttcaaacaaa gtttaaagct
     8761 aaagcacttc aagaaaaagt ttatattgaa tatgacaaag tgaaagcaga tagttgggat
     8821 agacgtaata tgcgtattga atttaatcca aacaaactta cacgagatga aatgatttgg
     8881 ttaaaacaaa atataataag ctacatggaa gatgacggtt ttacaagatt agatttagcc
     8941 tttgattttg aagatgattt gagtgactac tatgcaatgt ctgataaagc agttaagaaa
     9001 actatttttt atggtcgtaa tggtaagcca gaaacaaaat attttggcgt gagagatagt
     9061 aatagattta ttagaattta taataaaaag caagaacgta aagataatgc agatgctgaa
     9121 gttatgtctg aacatttatg gcgtgtagaa atcgaactta aaagagatat ggtggattac
     9181 tggaatgatt gctttagtga tttacatatc ttgcaaccag attggaaaac tatccaacgc
     9241 actgcggata gagcaatagt ttttatgtta ttgagtgatg aagaagaatg gggaaagctt
     9301 cacagaaatt ctagaacaaa atataagaat ttgataaaag aaatttcgcc agtcgattta
     9361 acggacttaa tgaaatcgac tttaaaagcg aacgaaaaac aattgcaaaa acaaatcgat
     9421 ttttggcaac atgaatttaa attttggaaa tagtgtacat attaatatta ctgaacaaaa
     9481 atgatatatt taaactattc taatttagga ggattttttt atgaagtgtc tatttaaaaa
     9541 tttggggaat ttatatgagg tgaaagaata atttacccct ataaacttta gtcacctcaa
     9601 gtaaagaggt aaaattgttt agtttatata aaaaatttaa aggtttgttt tatagcgttt
     9661 tattttggct ttgtattctt tcatttttta gtgtattaaa tgaaatggtt ttaaatgttt
     9721 ctttacctga tattgcaaat cattttaata ctactcctgg aattacaaac tgggtaaaca
     9781 ctgcatatat gttaactttt tcgataggaa cagcagtata tggaaaatta tctgattata
     9841 taaatataaa aaaattgtta attattggta ttagtttgag ctgtcttggt tcattgattg
     9901 cttttattgg gcccacctag gtattatcaa gataagaaag aaaaggattt ttcgctacgc
     9961 tcaaatcctt taaaaaaaca caaaagacca cattttttaa tgtggtcttt tattcttcaa
    10021 ctaaagcacc cattagttca acaaacgaaa attggataaa gtgggatatt tttaaaatat
    10081 atatttatgt tacagtaata ttgactttta aaaaaggatt gattctaatg aagaaagcag
    10141 acaagtaagc ctcctaaatt cactttagat aaaaatttag gaggcatatc aaatgaactt
    10201 taataaaatt gatttagaca attggaagag aaaagagata tttaatcatt atttgaacca
    10261 acaaacgact tttagtataa ccacagaaat tgatattagt gttttatacc gaaacataaa
    10321 acaagaagga tataaatttt accctgcatt tattttctta gtgacaaggg tgataaactc
    10381 aaatacagct tttagaactg gttacaatag cgacggagag ttaggttatt gggataagtt
    10441 agagccactt tatacaattt ttgatggtgt atctaaaaca ttctctggta tttggactcc
    10501 tgtaaagaat gacttcaaag agttttatga tttatacctt tctgatgtag agaaatataa
    10561 tggttcgggg aaattgtttc ccaaaacacc tatacctgaa aatgcttttt ctctttctat
    10621 tattccatgg acttcattta ctgggtttaa cttaaatatc aataataata gtaattacct
    10681 tctacccatt attacagcag gaaaattcat taataaaggt aattcaatat atttaccgct
    10741 atctttacag gtacatcatt ctgtttgtga tggttatcat gcaggattgt ttatgaactc
    10801 tattcaggaa ttgtcagata ggcctaatga ctggctttta taatatgaga taatgccgac
    10861 tgtacttttt acagtcggtt ttctaatgtc actaacctgc cccgttagtt gaagaaggcc
    10921 gcggcctcga gcggccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
    10981 gccct
//
