LOCUS       Exported               11911 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Integrate plasmid of Streptococcus pneumoniae, for chromosome 
            integration of IPTG-inducible dCas9sp.
ACCESSION   .
VERSION     .
KEYWORDS    pJWV102-PL-dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11911)
  AUTHORS   Liu X, Gallay C, Kjos M, Domenech A, Slager J, van Kessel SP, Knoops
            K, Sorg RA, Zhang JR, Veening JW
  TITLE     High-throughput CRISPRi phenotyping identifies new essential genes 
            in Streptococcus pneumoniae.
  JOURNAL   Mol Syst Biol. 2017 May 10;13(5):931. doi: 10.15252/msb.20167449.
  PUBMED    28490437
REFERENCE   2  (bases 1 to 11911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11911
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      289..360
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      376..403
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    500..519
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             555..566
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             573..4679
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(5876..5898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5984..6124
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6039..6058
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6139..6156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6310..6898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6390..6409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7069..7929)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7692..7711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7930..8034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8102..8120
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac
       61 cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg
      121 atcatggcga ccacacccgt cctgtggatc tatcgatgca tgcagccggt accatcacaa
      181 gcactttggg acgttctccc ttagtgcttt tttgatttct cataggccgg cctgttagtc
      241 atatggacac ttaagctcta gacggtgatc aacacgctag ccaggcatca aataaaacga
      301 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      361 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag aattcactag
      421 taattgtgag gcggataaca attctcactc attctacagt ttattcttga cattgcactg
      481 tccccctggt ataataacta attgtgagcg ctcacaatta agatccccgg ggcggccgcg
      541 aattcaaaag atctaaagag gagaaaggat ctatggataa gaaatactca ataggcttag
      601 ctatcggcac aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta
      661 aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg
      721 ctcttttatt tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta
      781 gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga
      841 tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag
      901 acaagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg
      961 agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg
     1021 atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga
     1081 ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac
     1141 aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag
     1201 cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc
     1261 ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc
     1321 ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata
     1381 cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt
     1441 ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta agagtaaata
     1501 ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc
     1561 aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa
     1621 tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag
     1681 aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat
     1741 tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta
     1801 ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt
     1861 atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt
     1921 attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg
     1981 aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat
     2041 catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa
     2101 aacatagttt gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg
     2161 ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg
     2221 atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca
     2281 aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt
     2341 cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg
     2401 aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg
     2461 agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac
     2521 agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta
     2581 ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca
     2641 atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa
     2701 aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta
     2761 gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag
     2821 taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa
     2881 ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag
     2941 aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa
     3001 agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata
     3061 ttaatcgttt aagtgattat gatgtcgatg ccattgttcc acaaagtttc cttaaagacg
     3121 attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg
     3181 ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca
     3241 agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg
     3301 aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc
     3361 atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta
     3421 ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt
     3481 tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa
     3541 atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct
     3601 atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag
     3661 gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa
     3721 ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg
     3781 gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc
     3841 cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa
     3901 ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa
     3961 aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg
     4021 aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg
     4081 aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag
     4141 ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc
     4201 gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa
     4261 gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag
     4321 aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta
     4381 ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag
     4441 ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta
     4501 ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa
     4561 caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc
     4621 aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgactaag
     4681 tcgacctcga gactagtcaa ggtcggcaat tctgcagtac taggacgccg ccaagccagc
     4741 ttaaacccag ctcaatgagc tgggtttttt gtttgttaaa aatgaagaag aaactgtgaa
     4801 gcgtatttat agcaaagcac tcaaaagttt accttatggg tgcttttttc gtgctttttt
     4861 gaaaagacaa aaaaaagaac cttgccaagc aagattcttt tgatagcgct atcgctgagc
     4921 gccggtcgct accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca
     4981 tgtctgcccg tatttcgcgt aaggaaatcc attatgtact atttctggtg atgaaatcaa
     5041 cgtaacattt aaagctgtca aagccaaagt catgagatgg cgtatggagc gtaaagctga
     5101 caagagcggt gttgcgatga ttgagatgac cttccttgca ccaagtgaat tgcctcaaga
     5161 aagcactcaa tcaaagattc ttgtagatgg aaaagaactt gctgatttcg ctgaaaatcg
     5221 tcaagactat caaattacct ataaaggtca acggccaaaa gtctcagttg aagaaaacaa
     5281 tcaagtagct tcaactgtgg tagatagtgg agaagatagc cttccagtac ttgttcgcct
     5341 cgtttcagaa agtggaaaac aagtcaagga ataccgtatc cagttgacta aggaaaaacc
     5401 agtttctgct gtacaagaag atcttccaaa actcgaattt gttgaaaaag atttggccta
     5461 caagacagtt gagaaaaaag attcaacact gtatctaggt gaaactcgtg tagaacaaga
     5521 aggaaaagtt ggaaaagaac gtatctttac agtgattaat cctgatggaa gtaaggaaga
     5581 aaaactccgt gaagtggtag aagttccgac agaccgcatc gtcttggttg gaaccaaacc
     5641 agtagctcaa gaagctaaaa aaccacaagt gtcagaaaaa gcagatacaa aaccaattga
     5701 ttcaagtgaa gctgatcaaa ctaataaagc ccagttacca aatacaggta gtgcggcaag
     5761 ccaagcagca gtagcagcag gtttagctct tctaggtttg agtgcaggat tagtagttac
     5821 taaaggtaaa aaagaagact agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     5881 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5941 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     6001 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     6061 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6121 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6721 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6841 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6901 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6961 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7021 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7081 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7141 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7201 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7261 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7321 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7381 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7441 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7501 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7561 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7621 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7681 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7741 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7801 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7861 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7921 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7981 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8041 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8101 aaataggcgt atcacgaggc cctttcgtct tcaagaatta attccttctt aacgccccaa
     8161 gttcatcacc aatgacatca actcacatga actacatgat gaacccagtt atcatggttt
     8221 tggataagat ttttgaaaaa ttcttcccag gccttgataa atatgacttt gatgctgcta
     8281 aattgaacaa gaaaatcggt ttctggggat ctaaattctt catcggtttc atccttggta
     8341 tcgttatcgg tattatggga actccacatc caattgcagg tgttgcagat gcagataaat
     8401 ggcgtcttgt tatcaaagga tggttgtctc ttggtttgac tgccggtgta tctttggaac
     8461 tcttctcact tatcggttca tggttcatcg cagccgtaga accactatca caaggtatta
     8521 caaacgttgc tactaaacgt cttcaaggac gtaaattcaa tatcggtctt gactggccat
     8581 tcatcgctgg tcgtgctgaa atctgggctt gtgccaacgt acttgcacca atcatgttga
     8641 ttgaagcagt gcttctttca aaagttggaa atggtatctt gccacttgca ggtatcatcg
     8701 ctatgggtgt tactccagct ctcttggttg taactcgtgg taaattgctc cgtatgatta
     8761 tcttcggaac actcttgttg ccactcttcc ttctttcagg tacacttatt gcaccatttg
     8821 caacagaact tgctaaaggt gtaggtgcct tcccagaagg tgtgagccaa actcaattga
     8881 ttactcactc tactcttgaa ggaccaatcg aaaaacttct tggttggaca attggtaaca
     8941 ctacaactgg tgatatcaaa gcaatccttg gtgcagtagc cttccttgta ttctatatcg
     9001 gtatctttgc ttggtacaga aaacaaatga tcaaacgtaa cgaagagtac gcagcaaaag
     9061 caaaataatg cgctccgcta gctttacaga caaagaacta tccttaatgg aagcactagc
     9121 aaaaggtatc aatgaagcaa gaaatattga aggctagtca gtaaaattca gataaacaaa
     9181 aagagccgat aagatgagag tttcatttct caaattatcg gctctgcgtc ttggcgtctg
     9241 gctctttgat tgttattata ttcatttttt gaacattaat taaagtttcg ttcttacatt
     9301 tggggcagta taaagggaaa tttttaagta tagtatccac tcgtattcgt agtcttgttt
     9361 tatttccaca aataggacac aatatccact tgtagtttat aataacaatc tcctcctttc
     9421 cactttaatt caaatctata ttaaagaata tttcatctta tttaataaga aaccatattt
     9481 atataacaac ataaaacgca ctaagttatt ttattgaaca tatatcttac tttatctatc
     9541 tgactattta gacgacgggt ctggcaaaca ggttcgccag tggtaacctg atatcctttt
     9601 agctctgcta aacaaacact aagcccattt gtaaaaaaag ttaaatcatt gcgataatct
     9661 tgaatacatc gagcaggaat ttctccaata ataatgacct cattattttt cagttgagta
     9721 tttacgatat ttgcacaata tttgggagca tcgttatatg cccgtgaaag atattcctgt
     9781 ggtgcataaa ctttaaaact aagatatggc tctaacaatt ctgttccagc ttttctaaag
     9841 gcttgctcca atacaatagg agcaagcatc cgaaaatctg ctggggtact aacagggcta
     9901 tagtataagc catacttaaa acagatttta cagtccgtca cattccaacc atacaatcct
     9961 tgttcacaac catagcgtat cccttccata actgcatttt gaaatgattg atttaagtat
    10021 ccaagagaaa ccgagctctc atactgcatt ccacttccca acggaagcgg tgatacagat
    10081 aaaccaatgg aagcccagaa aggatttggc ggcacttcga tgtgaatggt atattctgca
    10141 ttttttaacg gtctctccat ataaatgact gtaggctctt ttagttctat ctccacatga
    10201 tacttttctt gcaacagtgc actaatcact tccatttgta ctttccctaa gaaagaaagt
    10261 ataatttcat gtgtcgtaga atccacgtaa tatcgtagaa gcggatcact atctgagatt
    10321 tccaaaaggg catcaagcaa catttctctc tgttcaggtt tactcggttc aacagttgtt
    10381 tgtagtagag ggtgcggatt ttcaatcttt tttctctgtg gcaatagttt tgtatctcca
    10441 agaacactat ttaacttcaa aaactcattt tgcaaaataa caatttctcc agaataagct
    10501 ctatcaatct tacataattc accatttatt gaagtataca tttctgtaac ttttattttt
    10561 tctttttctg atactctaac cgaatctcgt aaatgtagta ctccactata aaggcgtata
    10621 tatgcaagac gttgtctttt ttttgtatat tcaattttga aaacatttcc gcaaagttca
    10681 gacggacctc gatgtgttga tgaataaaat ttattagtaa taacttctat aaggttatca
    10741 atccctatat tactttttgc acttccatga taaagaggga acagagaaca attctgaaat
    10801 cttatgcttt cctcttgttc gagttccaat gcttctaatg atttaccgga catatatttc
    10861 tctaaaaggt catcgtttcc ctctattacc gtatcccatt gttcagattc ggtaaagttc
    10921 gtcacacaca cattaggata cagttctacc ttctgtttga ttacaatttc ggcagaaagt
    10981 ttctctttaa tatcctgata aaccgttgat aaatcaattc cattttggtc aatcttattg
    11041 ataaaaaaga ttgtgggaat ccccattttc ctaagtgcat gaaataatat acgagtttgt
    11101 gcttgtacgc catcttttgc agaaatcagt agaattgccc catctaaaac tgataatgaa
    11161 cgatatactt ctgctaagaa atccatatgt cctggcgtgt ctatgatgtt caccttcgta
    11221 ttttcccact gaaaagaggt tattcctgtc tgaattgtaa ttcctctctg acgttctaaa
    11281 agcgtattat ccgtcctcgt tgtacctttg tccacgcttc ctaattctgt aatcgctcca
    11341 ctgttatata ataagctttc tgttaaggta gtttttcctg catcaacatg agctaaaact
    11401 ccaatattaa taattttcat gtgattttcc tccattcaaa agcccaaaag ggcataaaaa
    11461 tcccagtgat aaatactctt atcactggga tttttatgca taaccatagg catacaaagc
    11521 atacagatat tctccggata ctttagaatc acatgataaa ggtattctta aactgggtac
    11581 aaaaaactaa gccctcctaa aaaaggacat ccaattattt gttcccgcta tcaaattgac
    11641 agtttattta agaatacctt gccgcatatt tattaactcc ttttaaatag atacttaaat
    11701 tatagcacgt aagagcatat ttgtaaagga atctccaatt ttttatcaaa aagagtacat
    11761 gattacaaag tatctgtaat catgtaccaa tatttgttat tttacaatct tccaattatc
    11821 ttgtttttct aatatcaact caaattgcga gatttgggtt gcctttgttt cttgatctaa
    11881 atatttcaca gcgacattca cgattacttg g
//
