LOCUS       sequence-220869-       12508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chloramphenicol resistant derivative of pCas plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12508)
  AUTHORS   Swings T, Marciano DC, Atri B, Bosserman RE, Wang C, Leysen M, Bonte
            C, Schalck T, Furey I, Van den Bergh B, Verstraeten N, Christie PJ, 
            Herman C, Lichtarge O, Michiels J
  TITLE     CRISPR-FRT targets shared sites in a knock-out collection for 
            off-the-shelf genome editing.
  JOURNAL   Nat Commun. 2018 Jun 8;9(1):2231. doi: 10.1038/s41467-018-04651-5.
  PUBMED    29884781
REFERENCE   2  (bases 1 to 12508)
  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: "Nat 
            Commun."; date: "2018-06-8"; volume: "9(1):2231. doi"; pages: " 
            10.1038/s41467-018-04651-5"
COMMENT     Sequence Label: pCas-CmR(+)
FEATURES             Location/Qualifiers
     source          1..12508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(14..89)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    119..135
                     /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)."
     CDS             complement(404..1486)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1448..1467
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1487..1564)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             complement(1697..2647)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2695..2917
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        3675..3777
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3778..4437
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3844..3863)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_RNA        complement(4506..4584)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     CDS             4880..8986
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4880..8983
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9116..9994)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRH
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        10021..10305
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     10228..10247
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             10332..10340
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             10349..10765
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             10771..11556
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             11553..12233
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      12234..12478
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
ORIGIN
        1 gatctcaaaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta
       61 ttttaacttg ctatttctag ctctaaaacc tggtaacagg attagcagat gtgtgaaatt
      121 gttatccgct cacaattcca cacaacatac gagccggatg attaattgtc aacagctcat
      181 ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc cagaacggga
      241 gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc ataccacagc
      301 ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgatg ataagctgtc
      361 aaaccagatc aattcgcgct aacttacatt aattgcgttg cgctcactgc ccgctttcca
      421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      481 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
      541 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
      601 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
      661 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
      721 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
      781 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
      841 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
      901 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
      961 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     1021 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     1081 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     1141 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     1201 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     1261 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     1321 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     1381 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     1441 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     1501 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg
     1561 tgtcaacgta aatgcatgcc gctcgccttc catgggtatg gacagttttc cctttgatat
     1621 gtaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac
     1681 aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc
     1741 gttgtttttg cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac
     1801 tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg
     1861 tttttcttag tccgttatgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac
     1921 cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt
     1981 caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat
     2041 tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa
     2101 actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta
     2161 tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca
     2221 tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta
     2281 actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga
     2341 acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt
     2401 ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc
     2461 tactgatgtt catcatctga acgtattggt tataagtgaa cgataccgtc cgttctttcc
     2521 ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt
     2581 catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt
     2641 cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta
     2701 gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga
     2761 cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt
     2821 gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa
     2881 agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt
     2941 attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta
     3001 aaggcttaag tagcaccctc gcaagctcgg ttgcggccgc aatcgggcaa atcgctgaat
     3061 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
     3121 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
     3181 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
     3241 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
     3301 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
     3361 atgcacccag taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact
     3421 cacgttaagg gattttgccg ttacgcacca ccccgtcagt agctgaacag gagggacagc
     3481 tgatagaaac agaagccact ggagcacctc aaaaacacca tcatacacta aatcagtaag
     3541 ttggcagcat cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc
     3601 gcagaataaa taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg
     3661 cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     3721 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     3781 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     3841 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     3901 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     3961 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     4021 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     4081 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     4141 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     4201 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     4261 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     4321 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     4381 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     4441 ttttaaggca gttattggtg ccctgcaggg tttgcagtca gagtagaata gaagtatcaa
     4501 aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct tattttaact
     4561 tgctatgctg ttttgaatgg ttccaacaag attattttat aacttttata acaaataatc
     4621 aaggagaaat tcaaagaaat ttatcagcca taaaacaata cttaatacta tagaatgata
     4681 acaaaataaa ctacttttta aaagaatttt gtgttataat ctatttatta ttaagtattg
     4741 ggtaatattt tttgaagaga tattttgaaa aagaaaaatt aaagcatatt aaactaattt
     4801 cggaggtcat taaaactatt attgaaatca tcaaactcat tatggattta atttaaactt
     4861 tttattttag gaggcaaaaa tggataagaa atactcaata ggcttagata tcggcacaaa
     4921 tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt
     4981 tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga
     5041 cagtggagag acagcggaag cgactcgtct caaacggaca gctcgtagaa ggtatacacg
     5101 tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga
     5161 tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga
     5221 acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac
     5281 tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat
     5341 ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt
     5401 aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca
     5461 attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc
     5521 acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa
     5581 aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc
     5641 aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga
     5701 tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa
     5761 gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa
     5821 ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct
     5881 tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca
     5941 atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag aattttataa
     6001 atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa
     6061 tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat
     6121 tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa
     6181 agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc
     6241 attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac
     6301 cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg
     6361 catgacaaac tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct
     6421 ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat
     6481 gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa
     6541 aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg
     6601 ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta
     6661 ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga
     6721 tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga
     6781 aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg
     6841 ccgttatact ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca
     6901 atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat
     6961 gcagctgatc catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc
     7021 tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa
     7081 aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca
     7141 taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca
     7201 gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca
     7261 gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta
     7321 ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag
     7381 tgattatgat gtcgatcaca ttgttccaca aagtttcctt aaagacgatt caatagacaa
     7441 taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga
     7501 agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca
     7561 acgtaagttt gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc
     7621 tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat
     7681 tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa
     7741 agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa
     7801 agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg
     7861 aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa
     7921 agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc
     7981 aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa
     8041 tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg
     8101 ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat
     8161 tgtcaagaaa acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag
     8221 aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt
     8281 tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc
     8341 gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt
     8401 tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt
     8461 aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct
     8521 ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa
     8581 ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca
     8641 aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag
     8701 tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata
     8761 taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc atttatttac
     8821 gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa
     8881 acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg
     8941 tctttatgaa acacgcattg atttgagtca gctaggaggt gactgaagta tattttagat
     9001 gaagattatt tcttaatcta gacatgagcg gatacatatt tgaatgtatt tagaaaaata
     9061 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcatcg atttattatg
     9121 acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc
     9181 tggccccggt gcatttttta aatacccgcg agaagtagag ttgatcgtca aaaccaacat
     9241 tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga
     9301 tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg
     9361 acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt
     9421 ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat
     9481 ggagcgactc gttaatcgct tccatgtgcc gcagtaacaa ttgctcaagc agatttatcg
     9541 ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt
     9601 cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg
     9661 acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat
     9721 accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca
     9781 aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc
     9841 gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga
     9901 gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc
     9961 ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag
    10021 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
    10081 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
    10141 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
    10201 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
    10261 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
    10321 aattcgagct ctaaggaggt tataaaaaat ggatattaat actgaaactg agatcaagca
    10381 aaagcattca ctaaccccct ttcctgtttt cctaatcagc ccggcatttc gcgggcgata
    10441 ttttcacagc tatttcagga gttcagccat gaacgcttat tacattcagg atcgtcttga
    10501 ggctcagagc tgggcgcgtc actaccagca gctcgcccgt gaagagaaag aggcagaact
    10561 ggcagacgac atggaaaaag gcctgcccca gcacctgttt gaatcgctat gcatcgatca
    10621 tttgcaacgc cacggggcca gcaaaaaatc cattacccgt gcgtttgatg acgatgttga
    10681 gtttcaggag cgcatggcag aacacatccg gtacatggtt gaaaccattg ctcaccacca
    10741 ggttgatatt gattcagagg tataaaacga atgagtactg cactcgcaac gctggctggg
    10801 aagctggctg aacgtgtcgg catggattct gtcgacccac aggaactgat caccactctt
    10861 cgccagacgg catttaaagg tgatgccagc gatgcgcagt tcatcgcatt actgatcgtt
    10921 gccaaccagt acggccttaa tccgtggacg aaagaaattt acgcctttcc tgataagcag
    10981 aatggcatcg ttccggtggt gggcgttgat ggctggtccc gcatcatcaa tgaaaaccag
    11041 cagtttgatg gcatggactt tgagcaggac aatgaatcct gtacatgccg gatttaccgc
    11101 aaggaccgta atcatccgat ctgcgttacc gaatggatgg atgaatgccg ccgcgaacca
    11161 ttcaaaactc gcgaaggcag agaaatcacg gggccgtggc agtcgcatcc caaacggatg
    11221 ttacgtcata aagccatgat tcagtgtgcc cgtctggcct tcggatttgc tggtatctat
    11281 gacaaggatg aagccgagcg cattgtcgaa aatactgcat acactgcaga acgtcagccg
    11341 gaacgcgaca tcactccggt taacgatgaa accatgcagg agattaacac tctgctgatc
    11401 gccctggata aaacatggga tgacgactta ttgccgctct gttcccagat atttcgccgc
    11461 gacattcgtg catcgtcaga actgacacag gccgaagcag taaaagctct tggattcctg
    11521 aaacagaaag ccgcagagca gaaggtggca gcatgacacc ggacattatc ctgcagcgta
    11581 ccgggatcga tgtgagagct gtcgaacagg gggatgatgc gtggcacaaa ttacggctcg
    11641 gcgtcatcac cgcttcagaa gttcacaacg tgatagcaaa accccgctcc ggaaagaagt
    11701 ggcctgacat gaaaatgtcc tacttccaca ccctgcttgc tgaggtttgc accggtgtgg
    11761 ctccggaagt taacgctaaa gcactggcct ggggaaaaca gtacgagaac gacgccagaa
    11821 ccctgtttga attcacttcc ggcgtgaatg ttactgaatc cccgatcatc tatcgcgacg
    11881 aaagtatgcg taccgcctgc tctcccgatg gtttatgcag tgacggcaac ggccttgaac
    11941 tgaaatgccc gtttacctcc cgggatttca tgaagttccg gctcggtggt ttcgaggcca
    12001 taaagtcagc ttacatggcc caggtgcagt acagcatgtg ggtgacgcga aaaaatgcct
    12061 ggtactttgc caactatgac ccgcgtatga agcgtgaagg cctgcattat gtcgtgattg
    12121 agcgggatga aaagtacatg gcgagttttg acgagatcgt gccggagttc atcgaaaaaa
    12181 tggacgaggc actggctgaa attggttttg tatttgggga gcaatggcga tgacgcatcc
    12241 tcacgataat atccgggtag gcgcaatcac tttcgtctac tccgttacaa agcgaggctg
    12301 ggtatttccc ggcctttctg ttatccgaaa tccactgaaa gcacagcggc tggctgagga
    12361 gataaataat aaacgagggg ctgtatgcac aaagcatctt ctgttgagtt aagaacgagt
    12421 atcgagatgg cacatagcct tgctcaaatt ggaatcaggt ttgtgccaat accagtagaa
    12481 acagacgaag aatccatggg tatggaca
//
