LOCUS       sequence-366226-       15069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cr3 helper plasmid for recombination and Cas9 induction; harbors the
            Cas9 endonuclease and a species-specific recombination system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15069)
  AUTHORS   Teufel M, Klein CA, Mager M, Sobetzko P
  TITLE     A multifunctional system for genome editing and large-scale 
            interspecies gene transfer.
  JOURNAL   Nat Commun. 2022 Jun 14;13(1):3430. doi: 10.1038/s41467-022-30843-1.
  PUBMED    35701417
REFERENCE   2  (bases 1 to 15069)
  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: "2022-06-14"; volume: "13(1):3430. doi"; pages: " 
            10.1038/s41467-022-30843-1"
COMMENT     Sequence Label: cr3Dd
FEATURES             Location/Qualifiers
     source          1..15069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1471..1755)
                     /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     complement(1529..1548)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1782..2660
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      complement(2675..2746)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2828..2860)
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     CDS             complement(2861..6964)
                     /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(2861..6964)
                     /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"
     promoter        6999..7054
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7000..7018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7055..7089
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     CDS             7102..7728
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(7178..7197)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     oriT            7776..7884
                     /note="incP origin of transfer"
     primer_bind     complement(8105..8122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(8239..9054)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(9420..10271)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(10258..11097)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(11608..12579)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKYKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            13818..13905
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      14247..14641
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
ORIGIN
        1 cattgctccc caaaagtgaa ccctatttcc gccagtgcct catccatctt ttcgataaac
       61 tccggcacca tttcgtcaaa ctcggccata tatttttgat cccgctcaac gacgacgtgg
      121 tgaaggcctt cgcgcttcat gcgcggatcg tagtttgcaa agaaccaagc gccttttccg
      181 gtcacccaca tgctgtattg cacctgcgcc atatacgccg atttgatggc ttcaaaccca
      241 ccaaggcgga atttcatgaa gtcacgagag gtgaatgggc atttcaattc aaggccaaaa
      301 tcgttactgc acagtccatc tggggagcag gctgtgcgca tggtttcatc tcggaataag
      361 atcggcgact cagtgacagt tacgcctgta gtgaattcga acaacgagcg ggcgtcatct
      421 tcatactgct taccccacgc cagcgccttt gcattaactt ccggcgctac gccggtacaa
      481 acctcggcaa gaagggtgtg gaagtaggac attttcatgt ccgtccactt cgtgcctgat
      541 cgcggctttg aaatgacgtt atgtacctct gatgcggtaa tgacgccgag gcgcaggcga
      601 tgccacgcct catcaccttg ctcgatggtt ctcacgtcaa tcccagtgcg cgagtagatg
      661 gcttcagttg tcatcatgct gtcgcctttt gcttgaggaa tcccagcgct ttaaccgcct
      721 ctggctcagt cagatcagca ggctgcgtga tttctcgctt gaatattttt gagcacagcg
      781 gcaacaaatc gctgtcccat gatttgtcta acgtgacgag cagatcgttt atttcctgca
      841 tcgttgattc agtgactggc gtaatgtcgc gttccgcttg gcggtctgat gtgtaagttg
      901 tgctttcgac gatgcgctcg gcttcgtctt tgtcgtaaat accagcaaaa ccgaacgcta
      961 ggcgcgcgca ctgaatcatt gctttgtggc gcagcatacg tttcgggtgc gactgccagg
     1021 ggcctgtgat tttacgacca tcgcgcgttt cgaatggctc tctgcggcat tcatccatcc
     1081 actcagtaac gcagatcgga tggttgcgat ccttgcggta aatgcggcag gtgcatgatt
     1141 cgctatcctg ctcaaaatcc atgccatcaa actgctggtt ttcattgatg atacgtgacc
     1201 agccatcaac accaactact ggaacaatgc cgctttgctt gtcaggaaag gcgtaaatct
     1261 ccttcgtcca tggattaagt ccatactgat tggcgacgat cagcagggcg ataaactggg
     1321 cttcgctggc gtcccctttg aatgcggtgg cgcgcaacgt cgtgatcaat tcctgtgatt
     1381 caaccgaatt cattccgacg cgctcagcca attgaccagc caatgttgcg agagctgtgc
     1441 tcattcttgt ttttcctcat cgtcaaaatc atggagaaac agtagagagt tgcgataaaa
     1501 agcgtcaggt aggatccgct aatcttatgg ataaaaatgc tatggcatag caaagtgtga
     1561 cgccgtgcaa ataatcaatg tggacttttc tgccgtgatt atagacactt ttgttacgcg
     1621 tttttgtcat ggctttggtc ccgctttgtt acagaatgct tttaataagc ggggttaccg
     1681 gtttggttag cgagaagagc cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg
     1741 gacaattggt ttcttctctg aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg
     1801 atcccctgct gccgggatac tcgtttaatg cccatctggt ggcgggttta acgccgattg
     1861 aggccaacgg ttatctcgat ttttttatcg accgaccgct gggaatgaaa ggttatattc
     1921 tcaatctcac cattcgcggt cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc
     1981 gaccgggtga tattttgctg ttcccgccag gagagattca tcactacggt cgtcatccgg
     2041 aggctcgcga atggtatcac cagtgggttt actttcgtcc gcgcgcctac tggcatgaat
     2101 ggcttaactg gccgtcaata tttgccaata cggggttctt tcgcccggat gaagcgcacc
     2161 agccgcattt cagcgacctg tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct
     2221 attcggagct gctggcgata aatctgcttg agcaattgtt actgcggcgc atggaagcga
     2281 ttaacgagtc gctccatcca ccgatggata atcgggtacg cgaggcttgt cagtacatca
     2341 gcgatcacct ggcagacagc aattttgata tcgccagcgt cgcacagcat gtttgcttgt
     2401 cgccgtcgcg tctgtcacat cttttccgcc agcagttagg gattagcgtc ttaagctggc
     2461 gcgaggacca acgtatcagc caggcgaagc tgcttttgag caccacccgg atgcctatcg
     2521 ccaccgtcgg tcgcaatgtt ggttttgacg atcaactcta tttctcgcgg gtatttaaaa
     2581 aatgcaccgg ggccagcccg agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg
     2641 tagccgtcaa gttgtcataa tctcgactct agtagagagc gttcaccgac aaacaacaga
     2701 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggagatcc
     2761 ttactcgagc tccgtactca gagcgtagct gcgtccagcg gatgcctgga gatccttact
     2821 cgagttaagc agccagagcg tagttttcgt cgttagcagc gtcacctcct agctgactca
     2881 aatcaatgcg tgtttcataa agaccagtga tggattgatg gataagagtg gcatctaaaa
     2941 cttcttttgt agacgtatat cgtttacgat caattgttgt atcaaaatat ttaaaagcag
     3001 cgggagctcc aagattcgtc aacgtaaata aatgaataat attttctgct tgttcacgta
     3061 ttggtttgtc tctatgtttg ttatatgcac taagaacttt atctaaattg gcatctgcta
     3121 aaataacacg cttagaaaat tcactgattt gctcaataat ctcatctaaa taatgcttat
     3181 gctgctccac aaacaattgt ttttgttcgt tatcttctgg actacccttc aacttttcat
     3241 aatgactagc taaatataaa aaattcacat atttgcttgg cagagccagc tcatttcctt
     3301 tttgtaattc tccggcacta gccagcatcc gtttacgacc gttttctaac tcaaaaagac
     3361 tatatttagg tagtttaatg attaagtctt ttttaacttc cttatatcct ttagcttcta
     3421 aaaagtcaat cggatttttt tcaaaggaac ttctttccat aattgtgatc cctagtaact
     3481 ctttaacgga ttttaacttc ttcgatttcc ctttttccac cttagcaacc actaggactg
     3541 aataagctac cgttggacta tcaaaaccac catatttttt tggatcccag tcttttttac
     3601 gagcaataag cttgtccgaa tttctttttg gtaaaattga ctccttggag aatccgcctg
     3661 tctgtacttc tgttttcttg acaatattga cttggggcat ggacaatact ttgcgcactg
     3721 tggcaaaatc tcgcccttta tcccagacaa tttctccagt ttccccatta gtttcgatta
     3781 gagggcgttt gcgaatctct ccatttgcaa gtgtaatttc tgttttgaag aagttcatga
     3841 tattagagta aaagaaatat tttgcggttg ctttgcctat ttcttgctca gacttagcaa
     3901 tcattttacg aacatcataa actttataat caccatagac aaactccgat tcaagttttg
     3961 gatatttctt aatcaaagca gttccaacga cggcatttag atacgcatca tgggcatgat
     4021 ggtaattgtt aatctcacgt actttataga attggaaatc ttttcggaag tcagaaacta
     4081 atttagattt taaggtaatc actttaacct ctcgaataag tttatcattt tcatcgtatt
     4141 tagtattcat gcgactatcc aaaatttgtg ccacatgctt agtgatttgg cgagtttcaa
     4201 ccaattggcg tttgataaaa ccagctttat caagttcact caaacctcca cgttcagctt
     4261 tcgttaaatt atcaaactta cgttgagtga ttaacttggc gtttagaagt tgtctccaat
     4321 agtttttcat ctttttgact acttcttcac ttggaacgtt atccgattta ccacgatttt
     4381 tatcagaacg cgttaagacc ttattgtcta ttgaatcgtc tttaaggaaa ctttgtggaa
     4441 caatgtgatc gacatcataa tcacttaaac gattaatatc taattcttgg tccacataca
     4501 tgtctcttcc attttggaga taatagagat agagcttttc attttgcaat tgagtatttt
     4561 caacaggatg ctctttaaga atctgacttc ctaattcttt gataccttct tcgattcgtt
     4621 tcatacgctc tcgcgaattt ttctggccct tttgagttgt ctgattttca cgtgccattt
     4681 caataacgat attttctggc ttatgccgcc ccattacttt gaccaattca tcaacaactt
     4741 ttacagtctg taaaatacct tttttaatag cagggctacc agctaaattt gcaatatgtt
     4801 catgtaaact atcgccttgt ccagacactt gtgctttttg aatatcttct ttaaatgtca
     4861 aactatcatc atggatcagc tgcataaaat tgcgattggc aaaaccatct gatttcaaaa
     4921 aatctaatat tgttttgcca gattgcttat ccctaatacc attaatcaat tttcgagaca
     4981 aacgtcccca accagtataa cggcgacgtt taagctgttt catcacctta tcatcaaaga
     5041 ggtgagcata tgttttaagt ctttcctcaa tcatctccct atcttcaaat aaggtcaatg
     5101 ttaaaacaat atcctctaag atatcttcat tttcttcatt atccaaaaaa tctttatctt
     5161 taataatttt tagcaaatca tggtaggtac ctaatgaagc attaaatcta tcttcaactc
     5221 ctgaaatttc aacactatca aaacattcta tttttttgaa ataatcttct tttaattgct
     5281 taacggttac ttttcgattt gttttgaaga gtaaatcaac aatggctttc ttctgttcac
     5341 ctgaaagaaa tgctggtttt cgcattcctt cagtaacata tttgaccttt gtcaattcgt
     5401 tataaaccgt aaaatactca taaagcaaac tatgttttgg tagtactttt tcatttggaa
     5461 gatttttatc aaagtttgtc atgcgttcaa taaatgattg agctgaagca cctttatcga
     5521 caacttcttc aaaattccat ggggtaattg tttcttcaga cttccgagtc atccatgcaa
     5581 aacgactatt gccacgcgcc aatggaccaa cataataagg aattcgaaaa gtcaagattt
     5641 tttcaatctt ctcacgattg tcttttaaaa atggataaaa atcttcttgc cttctcaaaa
     5701 tagcatgcag ctcacccaag tgaatttgat ggggaataga gccgttgtca aaggtccgtt
     5761 gcttgcgcag caaatcttca cgatttagtt tcaccaataa ttcctcagta ccatccattt
     5821 tttctaaaat tggtttgata aatttataaa attcttcttg gctagctccc ccatcaatat
     5881 aacctgcata tccgtttttt gattgatcaa aaaagatttc tttatacttt tctggaagtt
     5941 gttgtcgaac taaagctttt aaaagagtca agtcttgatg atgttcatcg tagcgtttaa
     6001 tcattgaagc tgatagggga gccttagtta tttcagtatt tactcttagg atatctgaaa
     6061 gtaaaatagc atctgataaa ttcttagctg ccaaaaacaa atcagcatat tgatctccaa
     6121 tttgcgccaa taaattatct aaatcatcat cgtaagtatc ttttgaaagc tgtaatttag
     6181 catcttctgc caaatcaaaa tttgatttaa aattaggggt caaacccaat gacaaagcaa
     6241 tgagattccc aaataagcca tttttcttct caccggggag ctgagcaatg agattttcta
     6301 atcgtcttga tttactcaat cgtgcagaaa gaatcgcttt agcatctact ccacttgcgt
     6361 taatagggtt ttcttcaaat aattgattgt aggtttgtac caactggata aatagtttgt
     6421 ccacatcact attatcagga tttaaatctc cctcaatcaa aaaatgacca cgaaacttaa
     6481 tcatatgcgc taaggccaaa tagattaagc gcaaatccgc tttatcagta gaatctacca
     6541 atttttttcg cagatgatag atagttggat atttctcatg ataagcaact tcatctacta
     6601 tatttccaaa aataggatga cgttcatgct tcttatcttc ttccaccaaa aaagactctt
     6661 caagtcgatg aaagaaacta tcatctactt tcgccatctc atttgaaaaa atctcctgta
     6721 gataacaaat acgattcttc cgacgtgtat accttctacg agctgtccgt ttcagacgag
     6781 tcgcttccgc tgtctctcca ctgtcaaata aaagagcccc tataagattt tttttgatac
     6841 tgtggcggtc tgtatttccc agaaccttga actttttaga cggaacctta tattcatcag
     6901 tgatcaccgc ccatccgacg ctatttgtgc cgatatctaa gcctattgag tatttcttat
     6961 ccataagtgc gaccactctt tgaatctttt gaattctttt ctctatcact gatagggagt
     7021 ggtaaaataa ctctatcaac gatagagtgt caactttatg gctagctcag tcctaggtac
     7081 aatgctagca attaggaatt aatgatgtct agattagata aaagtaaagt gattaacagc
     7141 gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag
     7201 aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc
     7261 gacgccttag ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg
     7321 gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt
     7381 catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact
     7441 ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat
     7501 gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa
     7561 gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa
     7621 gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg
     7681 atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtcttaaga cttgttcaga
     7741 actcctggct cagagcgtag ctgcgtgcag cttatcggcc agcctcgcag agcaggattc
     7801 ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt
     7861 gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca
     7921 cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc
     7981 gctataatga ccccgaagca gggttatgca gcggaaagta taccttaagg agcgcgggca
     8041 aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc gttatgcagg
     8101 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     8161 actcaaaggc ggtaatctcg agtcccgtca agtcagcgta atgctctgcc agtgttacaa
     8221 ccaattaacc aattctgatt aaaagaactc gtccagcatc aggtgaaatt gcagtttgtt
     8281 catgtctggg ttatcgatgc cgtatttttg aaacagacgt ttttgcagag atgggctaaa
     8341 ttcacccaga cagttccaca gaattgccag atcttggtaa cgatctgcaa taccaacacg
     8401 accaacatca atacaaccga tcagtttacc ctcgtcaaaa atcaggttat ccagagaaaa
     8461 atcaccgtgg gtaacaacgc tatctggaga aaatggcagc agtttgtgca tttctttcca
     8521 aacttgttca actggccaac cgttacgttc atcatcaaaa tcagatgcat caaccagacc
     8581 gttgttcata cgagattgtg cttgtgccag acgaaaaaca cgatcgctgt taaatggaca
     8641 gttacaaact ggaatagagt gcagacgacg cagaaaaact gccagtgcat caacaatgtt
     8701 ttcaccagaa tctgggtatt cttccagaac ttgaaatgcg gttttacctg gaattgcggt
     8761 ggtcagcagc catgcatcat ctggggtacg aataaagtgt ttaatggttg gcagtggcat
     8821 aaattcggtc agccagttta aacgaaccat ttcatcggta acatcgtttg caacagaacc
     8881 tttaccgtgt ttcagaaaca gttctggtgc atctggttta ccgtaaagac ggtaaatggt
     8941 tgcaccagat tgaccaacgt tatcacgtgc ccatttgtaa ccgtacagat ctgcatccat
     9001 gttagagttc agacgtggac gagaacaaga ggtttcacgt tgaatgtgag acataacacc
     9061 ccttgtatta ctgtttatgt aaagtaatcg tagctaatta taagacgacc tccctcgagt
     9121 gtcaactccg ccttcagagc gtagctgcgg ctagcgaatg taagttagcg cgaattgcaa
     9181 gctggccgac gcgctgggct acgtcttgct ggcgttcggg agcagaagag catacatctg
     9241 gaagcaaagc caggaaagcg gcctatggag ctgtgcggca gcgctcagta ggcaattttt
     9301 caaaatattg ttaagccttt tctgagcatg gtatttttca tggtattacc aattagcagg
     9361 aaaataagcc attgaatata aaagataaaa atgtcttgtt tacaatagag tggggggggt
     9421 cagcctgccg ccttgggccg ggtgatgtcg tacttgcccg ccgcgaactc ggttaccgtc
     9481 cagcccagcg cgaccagctc cggcaacgcc tcgcgcaccc gctggcggcg cttgcgcatg
     9541 gtcgaaccac tggcctctga cggccagaca tagccgcaca aggtatctat ggaagccttg
     9601 ccggttttgc cggggtcgat ccagccacac agccgctggt gcagcaggcg ggcggtttcg
     9661 ctgtccagcg cccgcacctc gtccatgctg atgcgcacat gctggccgcc acccatgacg
     9721 gcctgcgcga tcaaggggtt cagggccacg tacaggcgcc cgtccgcctc gtcgctggcg
     9781 tactccgaca gcagccgaaa cccctgccgc ttgcggccat tctgggcgat gatggatacc
     9841 ttccaaaggc gctcgatgca gtcctgtatg tgcttgagcg ccccaccact atcgacctct
     9901 gccccgattt cctttgccag cgcccgatag ctacctttga ccaccatggc atcagcggtg
     9961 acggcctccc acttgggttc caggaacagc cggagctgcc gtccgccttc ggtcttgggt
    10021 tccgggccaa gcactaggcc attaggccca gccatggcca ccagcccttg caggatgcgc
    10081 agatcatcag cgcccagcgg ctccgggccg ctgaactcga tccgcttgcc gtcgccgtag
    10141 tcatacgtca cgtccagctt gctgcgcttg cgctcgcccc gcttgagggc acggaacagg
    10201 ccgggggcca gacagtgcgc cgggtcgtgc cggacgtggc tgaggctgtg cttgttctta
    10261 ggcttcacca cggggcaccc ccttgctctt gcgctgcctc tccagcacgg cgggcttgag
    10321 caccccgccg tcatgccgcc tgaaccaccg atcagcgaac ggtgcgccat agttggcctt
    10381 gctcacaccg aagcggacga agaaccggcg ctggtcgtcg tccacacccc attcctcggc
    10441 ctcggcgctg gtcatgctcg acaggtagga ctgccagcgg atgttatcga ccagtaccga
    10501 gctgccccgg ctggcctgct gctggtcgcc tgcgcccatc atggccgcgc ccttgctggc
    10561 atggtgcagg aacacgatag agcacccggt atcggcggcg atggcctcca tgcgaccgat
    10621 gacctgggcc atggggccgc tggcgttttc ttcctcgatg tggaaccggc gcagcgtgtc
    10681 cagcaccatc aggcggcggc cctcggcggc gcgcttgagg ccgtcgaacc actccggggc
    10741 catgatgttg ggcaggctgc cgatcagcgg ctggatcagc aggccgtcag ccacggcttg
    10801 ccgttcctcg gcgctgaggt gcgccccaag ggcgtgcagg cggtgatgaa tggcggtggg
    10861 cggatcttcg gcgggcaggt agatcaccgg gccggtgggc agttcgccca cctccagcag
    10921 atccggcccg cctgcaatct gtgcggccag ttgcagggcc agcatggatt taccggcacc
    10981 accgggcgac accagcgccc cgaccgtacc ggccaccatg ttgggcaaaa cgtagtccag
    11041 cggtggcggc gctgctgcga acgcctccag aatattgata ggcttatggg tagccattga
    11101 ttgcctcctt tgcaggcagt tggtggttag gcgctggcgg ggtcactacc cccgccctgc
    11161 gccgctctga gttcttccag gcactcgcgc agcgcctcgt attcgtcgtc ggtcagccag
    11221 aacttgcgct gacgcatccc tttggccttc atgcgctcgg catatcgcgc ttggcgtaca
    11281 gcgtcagggc tggccagcag gtcgccggtc tgcttgtcct tttggtcttt catatcagtc
    11341 accgagaaac ttgccggggc cgaaaggctt atcttcgcgg aacaaggaca aggtgcagcc
    11401 gtcaaggtta aggctggcca tatcagcgac tgaaaagcgg ccagcctcgg ccttgtttga
    11461 cgtataacca aagccaccgg gcaaccaata gcccttgtca cttttgatca ggtagaccga
    11521 ccctgaagcg cttttttcgt attccataaa acccccttct gtgcgtgagt actcatagta
    11581 taacaggcgt gagtaccaac gcaagcacta catgctgaaa tctggcccgc ccctgtccat
    11641 gcctcgctgg cggggtgccg gtgcccgtgc cagctcggcc cgcgcaagct ggacgctggg
    11701 cagacccatg accttgctga cggtgcgctc gatgtaatcc gcttcgtggc cgggcttgcg
    11761 ctctgccagc gctgggctgg cctcggccat ggccttgccg atttcctcgg cactgcggcc
    11821 ccggctggcc agcttctgcg cggcgataaa gtcgcacttg ctgaggtcat caccgaagcg
    11881 cttgaccagc ccggccatct cgctgcggta ctcgtccagc gccgtgcgcc ggtggcggct
    11941 aagctgccgc tcgggcagtt cgaggctggc cagcctgcgg gccttctcct gctgccgctg
    12001 ggcctgctcg atctgctggc cagcctgctg caccagcgcc gggccagcgg tggcggtctt
    12061 gcccttggat tcacgcagca gcacccacgg ctgataaccg gcgcgggtgg tgtgcttgta
    12121 cttgcggttg gtgaagcccg ccaagcggcc atagtggcgg ctgtcggcgc tggccgggtc
    12181 ggcgtcgtac tcgctggcca gcgtccgggc aatctgcccc cgaagttcac cgcctgcggc
    12241 gtcggccacc ttgacccatg cctgatagtt cttcgggctg gtttccacta ccagggcagg
    12301 ctcccggccc tcggctttca tgtcatccag gtcaaactcg ctgaggtcgt ccaccagcac
    12361 cagaccatgc cgctcctgct cggcgggcct gatatacacg tcattgccct gggcattcat
    12421 ccgcttgagc catggcgtgt tctggagcac ttcggcggct gaccattccc ggttcatcat
    12481 ctggccggtg gtggcgtccc tgacgccgat atcgaagcgc tcacagccca tggccttgag
    12541 ctgtcggcct atggcctgca aagtcctgtc gttcttcatc gggccaccaa gcgcagccag
    12601 atcgagccgt cctcggttgt cagtggcgtc aggtcgagca agagcaacga tgcgatcagc
    12661 agcaccaccg taggcatcat ggaagccagc atcacggtta gccatagctt ccagtgccac
    12721 ccccgcgacg cgctccgggc gctctgcgcg gcgctgctca cctcggcggc tacctcccgc
    12781 aactctttgg ccagctccac ccatgccgcc cctgtctggc gctgggcttt cagccactcc
    12841 gccgcctgcg cctcgctggc ctgctgggtc tggctcatga cctgccgggc ttcgtcggcc
    12901 agtgtcgcca tgctctgggc cagcggttcg atctgctccg ctaactcgtt gatgcctctg
    12961 gatttcttca ctctgtcgat tgcgttcatg gtctattgcc tcccggtatt cctgtaagtc
    13021 gatgatctgg gcgttggcgg tgtcgatgtt cagggccacg tctgcccggt cggtgcggat
    13081 gccccggcct tccatctcca ccacgttcgg ccccaggtga acaccgggca ggcgctcgat
    13141 gccctgcgcc tcaagtgttc tgtggtcaat gcgggcgtcg tggccagccc gctctaatgc
    13201 ccggttggca tggtcggccc atgcctcgcg ggtctgctca agccatgcct tgggcttgag
    13261 cgcttcggtc ttttgtgccc cgcccttctc cggggtcttg ccgttgtacc gcttgaacca
    13321 ctgagcggcg ggccgctcga tgccgtcatt gatccgctcg gagatcatca ggtggcagtg
    13381 cgggttctcg ccgccaccgg catggatggc cagcgtatac ggcaggcgct cggcaccggt
    13441 caggtgctgg gcgaactcgg acgccagcgc cttctgctgg tcgagggtca gctcgaccgg
    13501 cagggcaaat tcgacctcct tgaacagccg cccattggcg cgttcataca ggtcggcagc
    13561 atcccagtag tcggcgggcc gctcgacgaa ctccggcatg tgcccggatt cggcgtgcaa
    13621 gacttcatcc atgtcgcggg catacttgcc ttcgcgctgg atgtagtcgg ccttggccct
    13681 ggccgattgg ccgcccgacc tgctgccggt tttcgccgta aggtgataaa tcgccatgct
    13741 gcctcgctgt tgcttttgct tttcggctcc atgcaatggc cctcggagag cgcaccgccc
    13801 gaagggtggc cgttaggcca gtttctcgaa gagaaaccgg taagtgcgcc ctcccctaca
    13861 aagtagggtc gggattgccg ccgctgtgcc tccatgatag cctacgagac agcacattaa
    13921 caatggggtg tcaagatggt taaggggagc aacaaggcgg cggatcggct ggccaagctc
    13981 gaagaacaac gagcgcgaat caatgccgaa attcagcggg tgcgggcaag ggaacagcag
    14041 caagagcgca agaacgaaac aaggcgcaag gtgctggtgg gggccatgat tttggccaag
    14101 gtgaacagca gcgagtggcc ggaggatcgg ctcatggcgg caatggatgc gtaccttgaa
    14161 cgcgaccacg accgcgcctt gttcggtctg ccgccacgcc agaaggatga gccgggctga
    14221 atgatcgacc gagacaggcc ctgcggggct gcacacgcgc ccccaccctt cgggtagggg
    14281 gaaaggccgc taaagcggct aaaagcgctc cagcgtattt ctgcggggtt tggtgtgggg
    14341 tttagcgggc tttgcccgcc tttccccctg ccgcgcagcg gtggggcggt gtgtagccta
    14401 gcgcagcgaa tagaccagct atccggcctc tggccgggca tattgggcaa gggcagcagc
    14461 gccccacaag ggcgctgata accgcgccta gtggattatt cttagataat catggatgga
    14521 tttttccaac accccgccag cccccgcccc tgctgggttt gcaggtttgg gggcgtgaca
    14581 gttattgcag gggttcgtga cagttattgc aggggggcgt gacagttatt gcaggggttc
    14641 gtgacagtta gtacgggagt gacgggcact ggctggcaat gtctagcaac ggcaggcatt
    14701 tcggctgagg gtaaaagaac tttccgctaa gcgatagact gtatgtaaac acagtattgc
    14761 aaggacgcgg aacatgcctc atgtggcggc caggacggcc agccgggatc gggatactgg
    14821 tcgttaccag agccaccgac ccgagcaaac ccttctctat cagatcgttg acgagtatta
    14881 cccggcattc gctgcgctta tggcagagca gggaaaggaa ttgccgggct atgtgcaacg
    14941 ggaatttgaa gaatttctcc aatgcgggcg gctggagcat ggctttctac gggttcgctg
    15001 cgagtcttgc cacgccgagc acctggtcgc tctccgggct cagagcgtag ctgctgacag
    15061 cgtcatgtc
//
