LOCUS       sequence-363183-       14600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cr3 helper plasmid for recombination and Cas9 induction; harbors the
            Cas9 endonuclease and an V. natriegens-specific recombination 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14600)
  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 14600)
  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: cr3Vn
FEATURES             Location/Qualifiers
     source          1..14600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      349..743
                     /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)"
     oriT            1085..1172
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2411..3382
                     /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
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             3893..4732
                     /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             4719..5570
                     /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             6028..6843
                     /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"
     primer_bind     6960..6977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(7156..7782)
                     /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="MMSRLDKSRVISSALELLNEVGIEGLTTRKLAQRLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     7687..7706
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        7798..7853
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7804..7822
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    7834..7852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             7902..12005
                     /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             7902..12005
                     /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             12012..12044
                     /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"
     terminator      12062..12133
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             12560..12976
                     /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             12982..13767
                     /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             13764..14447
                     /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
                     EQWRA"
     terminator      14456..14527
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tagaaagcca tgctccagcc gcccgcattg gagaaattct tcaaattccc gttgcacata
       61 gcccggcaat tcctttccct gctctgccat aagcgcagcg aatgccgggt aatactcgtc
      121 aacgatctga tagagaaggg tttgctcggg tcggtggctc tggtaacgac cagtatcccg
      181 atcccggctg gccgtcctgg ccgccacatg aggcatgttc cgcgtccttg caatactgtg
      241 tttacataca gtctatcgct tagcggaaag ttcttttacc ctcagccgaa atgcctgccg
      301 ttgctagaca ttgccagcca gtgcccgtca ctcccgtact aactgtcacg aacccctgca
      361 ataactgtca cgcccccctg caataactgt cacgaacccc tgcaataact gtcacgcccc
      421 caaacctgca aacccagcag gggcgggggc tggcggggtg ttggaaaaat ccatccatga
      481 ttatctaaga ataatccact aggcgcggtt atcagcgccc ttgtggggcg ctgctgccct
      541 tgcccaatat gcccggccag aggccggata gctggtctat tcgctgcgct aggctacaca
      601 ccgccccacc gctgcgcggc agggggaaag gcgggcaaag cccgctaaac cccacaccaa
      661 accccgcaga aatacgctgg agcgctttta gccgctttag cggcctttcc ccctacccga
      721 agggtggggg cgcgtgtgca gccccgcagg gcctgtctcg gtcgatcatt cagcccggct
      781 catccttctg gcgtggcggc agaccgaaca aggcgcggtc gtggtcgcgt tcaaggtacg
      841 catccattgc cgccatgagc cgatcctccg gccactcgct gctgttcacc ttggccaaaa
      901 tcatggcccc caccagcacc ttgcgccttg tttcgttctt gcgctcttgc tgctgttccc
      961 ttgcccgcac ccgctgaatt tcggcattga ttcgcgctcg ttgttcttcg agcttggcca
     1021 gccgatccgc cgccttgttg ctccccttaa ccatcttgac accccattgt taatgtgctg
     1081 tctcgtaggc tatcatggag gcacagcggc ggcaatcccg accctacttt gtaggggagg
     1141 gcgcacttac cggtttctct tcgagaaact ggcctaacgg ccacccttcg ggcggtgcgc
     1201 tctccgaggg ccattgcatg gagccgaaaa gcaaaagcaa cagcgaggca gcatggcgat
     1261 ttatcacctt acggcgaaaa ccggcagcag gtcgggcggc caatcggcca gggccaaggc
     1321 cgactacatc cagcgcgaag gcaagtatgc ccgcgacatg gatgaagtct tgcacgccga
     1381 atccgggcac atgccggagt tcgtcgagcg gcccgccgac tactgggatg ctgccgacct
     1441 gtatgaacgc gccaatgggc ggctgttcaa ggaggtcgaa tttgccctgc cggtcgagct
     1501 gaccctcgac cagcagaagg cgctggcgtc cgagttcgcc cagcacctga ccggtgccga
     1561 gcgcctgccg tatacgctgg ccatccatgc cggtggcggc gagaacccgc actgccacct
     1621 gatgatctcc gagcggatca atgacggcat cgagcggccc gccgctcagt ggttcaagcg
     1681 gtacaacggc aagaccccgg agaagggcgg ggcacaaaag accgaagcgc tcaagcccaa
     1741 ggcatggctt gagcagaccc gcgaggcatg ggccgaccat gccaaccggg cattagagcg
     1801 ggctggccac gacgcccgca ttgaccacag aacacttgag gcgcagggca tcgagcgcct
     1861 gcccggtgtt cacctggggc cgaacgtggt ggagatggaa ggccggggca tccgcaccga
     1921 ccgggcagac gtggccctga acatcgacac cgccaacgcc cagatcatcg acttacagga
     1981 ataccgggag gcaatagacc atgaacgcaa tcgacagagt gaagaaatcc agaggcatca
     2041 acgagttagc ggagcagatc gaaccgctgg cccagagcat ggcgacactg gccgacgaag
     2101 cccggcaggt catgagccag acccagcagg ccagcgaggc gcaggcggcg gagtggctga
     2161 aagcccagcg ccagacaggg gcggcatggg tggagctggc caaagagttg cgggaggtag
     2221 ccgccgaggt gagcagcgcc gcgcagagcg cccggagcgc gtcgcggggg tggcactgga
     2281 agctatggct aaccgtgatg ctggcttcca tgatgcctac ggtggtgctg ctgatcgcat
     2341 cgttgctctt gctcgacctg acgccactga caaccgagga cggctcgatc tggctgcgct
     2401 tggtggcccg atgaagaacg acaggacttt gcaggccata ggccgacagc tcaaggccat
     2461 gggctgtgag cgcttcgata tcggcgtcag ggacgccacc accggccaga tgatgaaccg
     2521 ggaatggtca gccgccgaag tgctccagaa cacgccatgg ctcaagcgga tgaatgccca
     2581 gggcaatgac gtgtatatca ggcccgccga gcaggagcgg catggtctgg tgctggtgga
     2641 cgacctcagc gagtttgacc tggatgacat gaaagccgag ggccgggagc ctgccctggt
     2701 agtggaaacc agcccgaaga actatcaggc atgggtcaag gtggccgacg ccgcaggcgg
     2761 tgaacttcgg gggcagattg cccggacgct ggccagcgag tacgacgccg acccggccag
     2821 cgccgacagc cgccactatg gccgcttggc gggcttcacc aaccgcaagg acaagcacac
     2881 cacccgcgcc ggttatcagc cgtgggtgct gctgcgtgaa tccaagggca agaccgccac
     2941 cgctggcccg gcgctggtgc agcaggctgg ccagcagatc gagcaggccc agcggcagca
     3001 ggagaaggcc cgcaggctgg ccagcctcga actgcccgag cggcagctta gccgccaccg
     3061 gcgcacggcg ctggacgagt accgcagcga gatggccggg ctggtcaagc gcttcggtga
     3121 tgacctcagc aagtgcgact ttatcgccgc gcagaagctg gccagccggg gccgcagtgc
     3181 cgaggaaatc ggcaaggcca tggccgaggc cagcccagcg ctggcagagc gcaagcccgg
     3241 ccacgaagcg gattacatcg agcgcaccgt cagcaaggtc atgggtctgc ccagcgtcca
     3301 gcttgcgcgg gccgagctgg cacgggcacc ggcaccccgc cagcgaggca tggacagggg
     3361 cgggccagat ttcagcatgt agtgcttgcg ttggtactca cgcctgttat actatgagta
     3421 ctcacgcaca gaagggggtt ttatggaata cgaaaaaagc gcttcagggt cggtctacct
     3481 gatcaaaagt gacaagggct attggttgcc cggtggcttt ggttatacgt caaacaaggc
     3541 cgaggctggc cgcttttcag tcgctgatat ggccagcctt aaccttgacg gctgcacctt
     3601 gtccttgttc cgcgaagata agcctttcgg ccccggcaag tttctcggtg actgatatga
     3661 aagaccaaaa ggacaagcag accggcgacc tgctggccag ccctgacgct gtacgccaag
     3721 cgcgatatgc cgagcgcatg aaggccaaag ggatgcgtca gcgcaagttc tggctgaccg
     3781 acgacgaata cgaggcgctg cgcgagtgcc tggaagaact cagagcggcg cagggcgggg
     3841 gtagtgaccc cgccagcgcc taaccaccaa ctgcctgcaa aggaggcaat caatggctac
     3901 ccataagcct atcaatattc tggaggcgtt cgcagcagcg ccgccaccgc tggactacgt
     3961 tttgcccaac atggtggccg gtacggtcgg ggcgctggtg tcgcccggtg gtgccggtaa
     4021 atccatgctg gccctgcaac tggccgcaca gattgcaggc gggccggatc tgctggaggt
     4081 gggcgaactg cccaccggcc cggtgatcta cctgcccgcc gaagatccgc ccaccgccat
     4141 tcatcaccgc ctgcacgccc ttggggcgca cctcagcgcc gaggaacggc aagccgtggc
     4201 tgacggcctg ctgatccagc cgctgatcgg cagcctgccc aacatcatgg ccccggagtg
     4261 gttcgacggc ctcaagcgcg ccgccgaggg ccgccgcctg atggtgctgg acacgctgcg
     4321 ccggttccac atcgaggaag aaaacgccag cggccccatg gcccaggtca tcggtcgcat
     4381 ggaggccatc gccgccgata ccgggtgctc tatcgtgttc ctgcaccatg ccagcaaggg
     4441 cgcggccatg atgggcgcag gcgaccagca gcaggccagc cggggcagct cggtactggt
     4501 cgataacatc cgctggcagt cctacctgtc gagcatgacc agcgccgagg ccgaggaatg
     4561 gggtgtggac gacgaccagc gccggttctt cgtccgcttc ggtgtgagca aggccaacta
     4621 tggcgcaccg ttcgctgatc ggtggttcag gcggcatgac ggcggggtgc tcaagcccgc
     4681 cgtgctggag aggcagcgca agagcaaggg ggtgccccgt ggtgaagcct aagaacaagc
     4741 acagcctcag ccacgtccgg cacgacccgg cgcactgtct ggcccccggc ctgttccgtg
     4801 ccctcaagcg gggcgagcgc aagcgcagca agctggacgt gacgtatgac tacggcgacg
     4861 gcaagcggat cgagttcagc ggcccggagc cgctgggcgc tgatgatctg cgcatcctgc
     4921 aagggctggt ggccatggct gggcctaatg gcctagtgct tggcccggaa cccaagaccg
     4981 aaggcggacg gcagctccgg ctgttcctgg aacccaagtg ggaggccgtc accgctgatg
     5041 ccatggtggt caaaggtagc tatcgggcgc tggcaaagga aatcggggca gaggtcgata
     5101 gtggtggggc gctcaagcac atacaggact gcatcgagcg cctttggaag gtatccatca
     5161 tcgcccagaa tggccgcaag cggcaggggt ttcggctgct gtcggagtac gccagcgacg
     5221 aggcggacgg gcgcctgtac gtggccctga accccttgat cgcgcaggcc gtcatgggtg
     5281 gcggccagca tgtgcgcatc agcatggacg aggtgcgggc gctggacagc gaaaccgccc
     5341 gcctgctgca ccagcggctg tgtggctgga tcgaccccgg caaaaccggc aaggcttcca
     5401 tagatacctt gtgcggctat gtctggccgt cagaggccag tggttcgacc atgcgcaagc
     5461 gccgccagcg ggtgcgcgag gcgttgccgg agctggtcgc gctgggctgg acggtaaccg
     5521 agttcgcggc gggcaagtac gacatcaccc ggcccaaggc ggcaggctga ccccccccac
     5581 tctattgtaa acaagacatt tttatctttt atattcaatg gcttattttc ctgctaattg
     5641 gtaataccat gaaaaatacc atgctcagaa aaggcttaac aatattttga aaaattgcct
     5701 actgagcgct gccgcacagc tccataggcc gctttcctgg ctttgcttcc agatgtatgc
     5761 tcttctgctc ccgaacgcca gcaagacgta gcccagcgcg tcggccagct tgcaattcgc
     5821 gctaacttac attcgctagc cgcagctacg ctctgaaggc ggaggttaag atgcgtgatc
     5881 tgatccttca actcagcaaa agttcgattt attcaacaaa gccacgttgt gtctcaaaat
     5941 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     6001 tacataaaca gtaatacaag gggtgttatg tctcacattc aacgtgaaac ctcttgttct
     6061 cgtccacgtc tgaactctaa catggatgca gatctgtacg gttacaaatg ggcacgtgat
     6121 aacgttggtc aatctggtgc aaccatttac cgtctttacg gtaaaccaga tgcaccagaa
     6181 ctgtttctga aacacggtaa aggttctgtt gcaaacgatg ttaccgatga aatggttcgt
     6241 ttaaactggc tgaccgaatt tatgccactg ccaaccatta aacactttat tcgtacccca
     6301 gatgatgcat ggctgctgac caccgcaatt ccaggtaaaa ccgcatttca agttctggaa
     6361 gaatacccag attctggtga aaacattgtt gatgcactgg cagtttttct gcgtcgtctg
     6421 cactctattc cagtttgtaa ctgtccattt aacagcgatc gtgtttttcg tctggcacaa
     6481 gcacaatctc gtatgaacaa cggtctggtt gatgcatctg attttgatga tgaacgtaac
     6541 ggttggccag ttgaacaagt ttggaaagaa atgcacaaac tgctgccatt ttctccagat
     6601 agcgttgtta cccacggtga tttttctctg gataacctga tttttgacga gggtaaactg
     6661 atcggttgta ttgatgttgg tcgtgttggt attgcagatc gttaccaaga tctggcaatt
     6721 ctgtggaact gtctgggtga atttagccca tctctgcaaa aacgtctgtt tcaaaaatac
     6781 ggcatcgata acccagacat gaacaaactg caatttcacc tgatgctgga cgagttcttt
     6841 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     6901 tcgagattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6961 gcgagtcagt gagcgaggaa gcctgcataa cgcgaagtaa tcttttcggt tttaaagaaa
     7021 aagggcaggg tggtgacacc ttgcccgcgc tgcacgcagc tacgctctga gccaggagtt
     7081 ttgttatcaa taaaaaaggc cccccgttag ggaggcttat tgttcgtccc atcacggaaa
     7141 aaggttatgc tgcttttaag acccactttc acatttaagt tgtttttcta atccgcatat
     7201 gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgatcaa ataattcgat
     7261 agcttgtcgt aataatggcg gcatactatc agtagtaggt gtttcccttt cttctttagc
     7321 gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca cagcgctgag
     7381 tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta attgattttc
     7441 gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg
     7501 atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat cttgccagct
     7561 ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa tggctaaggc
     7621 gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct ctacacctag
     7681 cctctgggcg agtttacggg ttgttaaacc ttcgattccg acctcattaa gcagctctaa
     7741 tgcgctgcta atcactctac ttttatctaa tcgagacatc attaattcct aatttttgtt
     7801 gacactctat cattgataga gttattttac cactccctat cagtgataga gaaaagtgaa
     7861 tactaaagaa ttcaaaagat ctaaagagga cttcggatct aatggataag aaatactcaa
     7921 taggcttaga tatcggcaca aatagcgtcg gatgggcggt gatcactgat gaatataagg
     7981 ttccgtctaa aaagttcaag gttctgggaa atacagaccg ccacagtatc aaaaaaaatc
     8041 ttataggggc tcttttattt gacagtggag agacagcgga agcgactcgt ctgaaacgga
     8101 cagctcgtag aaggtataca cgtcggaaga atcgtatttg ttatctacag gagatttttt
     8161 caaatgagat ggcgaaagta gatgatagtt tctttcatcg acttgaagag tcttttttgg
     8221 tggaagaaga taagaagcat gaacgtcatc ctatttttgg aaatatagta gatgaagttg
     8281 cttatcatga gaaatatcca actatctatc atctgcgaaa aaaattggta gattctactg
     8341 ataaagcgga tttgcgctta atctatttgg ccttagcgca tatgattaag tttcgtggtc
     8401 attttttgat tgagggagat ttaaatcctg ataatagtga tgtggacaaa ctatttatcc
     8461 agttggtaca aacctacaat caattatttg aagaaaaccc tattaacgca agtggagtag
     8521 atgctaaagc gattctttct gcacgattga gtaaatcaag acgattagaa aatctcattg
     8581 ctcagctccc cggtgagaag aaaaatggct tatttgggaa tctcattgct ttgtcattgg
     8641 gtttgacccc taattttaaa tcaaattttg atttggcaga agatgctaaa ttacagcttt
     8701 caaaagatac ttacgatgat gatttagata atttattggc gcaaattgga gatcaatatg
     8761 ctgatttgtt tttggcagct aagaatttat cagatgctat tttactttca gatatcctaa
     8821 gagtaaatac tgaaataact aaggctcccc tatcagcttc aatgattaaa cgctacgatg
     8881 aacatcatca agacttgact cttttaaaag ctttagttcg acaacaactt ccagaaaagt
     8941 ataaagaaat cttttttgat caatcaaaaa acggatatgc aggttatatt gatgggggag
     9001 ctagccaaga agaattttat aaatttatca aaccaatttt agaaaaaatg gatggtactg
     9061 aggaattatt ggtgaaacta aatcgtgaag atttgctgcg caagcaacgg acctttgaca
     9121 acggctctat tccccatcaa attcacttgg gtgagctgca tgctattttg agaaggcaag
     9181 aagattttta tccattttta aaagacaatc gtgagaagat tgaaaaaatc ttgacttttc
     9241 gaattcctta ttatgttggt ccattggcgc gtggcaatag tcgttttgca tggatgactc
     9301 ggaagtctga agaaacaatt accccatgga attttgaaga agttgtcgat aaaggtgctt
     9361 cagctcaatc atttattgaa cgcatgacaa actttgataa aaatcttcca aatgaaaaag
     9421 tactaccaaa acatagtttg ctttatgagt attttacggt ttataacgaa ttgacaaagg
     9481 tcaaatatgt tactgaagga atgcgaaaac cagcatttct ttcaggtgaa cagaagaaag
     9541 ccattgttga tttactcttc aaaacaaatc gaaaagtaac cgttaagcaa ttaaaagaag
     9601 attatttcaa aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt gaagatagat
     9661 ttaatgcttc attaggtacc taccatgatt tgctaaaaat tattaaagat aaagattttt
     9721 tggataatga agaaaatgaa gatatcttag aggatattgt tttaacattg accttatttg
     9781 aagataggga gatgattgag gaaagactta aaacatatgc tcacctcttt gatgataagg
     9841 tgatgaaaca gcttaaacgt cgccgttata ctggttgggg acgtttgtct cgaaaattga
     9901 ttaatggtat tagggataag caatctggca aaacaatatt agattttttg aaatcagatg
     9961 gttttgccaa tcgcaatttt atgcagctga tccatgatga tagtttgaca tttaaagaag
    10021 atattcaaaa agcacaagtg tctggacaag gcgatagttt acatgaacat attgcaaatt
    10081 tagctggtag ccctgctatt aaaaaaggta ttttacagac tgtaaaagtt gttgatgaat
    10141 tggtcaaagt aatggggcgg cataagccag aaaatatcgt tattgaaatg gcacgtgaaa
    10201 atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg tatgaaacga atcgaagaag
    10261 gtatcaaaga attaggaagt cagattctta aagagcatcc tgttgaaaat actcaattgc
    10321 aaaatgaaaa gctctatctc tattatctcc aaaatggaag agacatgtat gtggaccaag
    10381 aattagatat taatcgttta agtgattatg atgtcgatca cattgttcca caaagtttcc
    10441 ttaaagacga ttcaatagac aataaggtct taacgcgttc tgataaaaat cgtggtaaat
    10501 cggataacgt tccaagtgaa gaagtagtca aaaagatgaa aaactattgg agacaacttc
    10561 taaacgccaa gttaatcact caacgtaagt ttgataattt aacgaaagct gaacgtggag
    10621 gtttgagtga acttgataaa gctggtttta tcaaacgcca attggttgaa actcgccaaa
    10681 tcactaagca tgtggcacaa attttggata gtcgcatgaa tactaaatac gatgaaaatg
    10741 ataaacttat tcgagaggtt aaagtgatta ccttaaaatc taaattagtt tctgacttcc
    10801 gaaaagattt ccaattctat aaagtacgtg agattaacaa ttaccatcat gcccatgatg
    10861 cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa cttgaatcgg
    10921 agtttgtcta tggtgattat aaagtttatg atgttcgtaa aatgattgct aagtctgagc
    10981 aagaaatagg caaagcaacc gcaaaatatt tcttttactc taatatcatg aacttcttca
    11041 aaacagaaat tacacttgca aatggagaga ttcgcaaacg ccctctaatc gaaactaatg
    11101 gggaaactgg agaaattgtc tgggataaag ggcgagattt tgccacagtg cgcaaagtat
    11161 tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt acagacaggc ggattctcca
    11221 aggagtcaat tttaccaaaa agaaattcgg acaagcttat tgctcgtaaa aaagactggg
    11281 atccaaaaaa atatggtggt tttgatagtc caacggtagc ttattcagtc ctagtggttg
    11341 ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt taaagagtta ctagggatca
    11401 caattatgga aagaagttcc tttgaaaaaa atccgattga ctttttagaa gctaaaggat
    11461 ataaggaagt taaaaaagac ttaatcatta aactacctaa atatagtctt tttgagttag
    11521 aaaacggtcg taaacggatg ctggctagtg ccggagaatt acaaaaagga aatgagctgg
    11581 ctctgccaag caaatatgtg aattttttat atttagctag tcattatgaa aagttgaagg
    11641 gtagtccaga agataacgaa caaaaacaat tgtttgtgga gcagcataag cattatttag
    11701 atgagattat tgagcaaatc agtgaatttt ctaagcgtgt tattttagca gatgccaatt
    11761 tagataaagt tcttagtgca tataacaaac atagagacaa accaatacgt gaacaagcag
    11821 aaaatattat tcatttattt acgttgacga atcttggagc tcccgctgct tttaaatatt
    11881 ttgatacaac aattgatcgt aaacgatata cgtctacaaa agaagtttta gatgccactc
    11941 ttatccatca atccatcact ggtctttatg aaacacgcat tgatttgagt cagctaggag
    12001 gtgacgcttt agctgctaac gacgaaaact acgctctggc tgcttagggg taaccaggca
    12061 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
    12121 ggtgaacgct ctccgctgga cgcagctacg ctctgagtac ggagtgttat ccatccactg
    12181 gtagaggtga gtgttcgcta tacatatttg tggatttagg gacatttgtt agtgacaaaa
    12241 atcacagcgg aaaaatgtag cgaatttgtc cattcattta gccagtgtgg ctatgacaca
    12301 gatctcaatt atgcgaccaa tgatccaaat tctcagtaag caacccaata ccagcctaat
    12361 gcaaacgttg aattcgctgg tttcttgctt ttccgacctg acagaagaag gtgttaaaaa
    12421 gaacacaaaa aatcgtccat gacgtttttg tccataaagt tagcagacct ctttatggga
    12481 taacatccct cctctagcta taacaacaag tagatttagt ttgctgacca aaatactaga
    12541 gaaagagggg aaataatcaa tggatattaa tactgaaact gagatcaagc aaaagcattc
    12601 actaaccccc tttcctgttt tcctaatcag cccggcattt cgcgggcgat attttcacag
    12661 ctatttcagg agttcagcca tgaacgctta ttacattcag gatcgtcttg aggctcagag
    12721 ctgggcgcgt cactaccagc agctcgcccg tgaagagaaa gaggcagaac tggcagacga
    12781 catggaaaaa ggcctgcccc agcacctgtt tgaatcgcta tgcatcgatc atttgcaacg
    12841 ccacggggcc agcaaaaaat ccattacccg tgcgtttgat gacgatgttg agtttcagga
    12901 gcgcatggca gaacacatcc ggtacatggt tgaaaccatt gctcaccacc aggttgatat
    12961 tgattcagag gtataaaacg aatgagtact gcactcgcaa cgctggctgg gaagctggct
    13021 gaacgtgtcg gcatggattc tgtcgaccca caggaactga tcaccactct tcgccagacg
    13081 gcatttaaag gtgatgccag cgatgcgcag ttcatcgcat tactgatcgt tgccaaccag
    13141 tacggcctta atccgtggac gaaagaaatt tacgcctttc ctgataagca gaatggcatc
    13201 gttccggtgg tgggcgttga tggctggtcc cgcatcatca atgaaaacca gcagtttgat
    13261 ggcatggact ttgagcagga caatgaatcc tgtacatgcc ggatttaccg caaggaccgt
    13321 aatcatccga tctgcgttac cgaatggatg gatgaatgcc gccgcgaacc attcaaaact
    13381 cgcgaaggca gagaaatcac ggggccgtgg cagtcgcatc ccaaacggat gttacgtcat
    13441 aaagccatga ttcagtgtgc ccgtctggcc ttcggatttg ctggtatcta tgacaaggat
    13501 gaagccgagc gcattgtcga aaatactgca tacactgcag aacgtcagcc ggaacgcgac
    13561 atcactccgg ttaacgatga aaccatgcag gagattaaca ctctgctgat cgccctggat
    13621 aaaacatggg atgacgactt attgccgctc tgttcccaga tatttcgccg cgacattcgt
    13681 gcatcgtcag aactgacaca ggccgaagca gtaaaagctc ttggattcct gaaacagaaa
    13741 gccgcagagc agaaggtggc agcatgacac cggacattat cctgcagcgt accgggatcg
    13801 atgtgagagc tgtcgaacag ggggatgatg cgtggcacaa attacggctc ggcgtcatca
    13861 ccgcttcaga agttcacaac gtgatagcaa aaccccgctc cggaaagaag tggcctgaca
    13921 tgaaaatgtc ctacttccac accctgcttg ctgaggtttg caccggtgtg gctccggaag
    13981 ttaacgctaa agcactggcc tggggaaaac agtacgagaa cgacgccaga accctgtttg
    14041 aattcacttc cggcgtgaat gttactgaat ccccgatcat ctatcgcgac gaaagtatgc
    14101 gtaccgcctg ctctcccgat ggtttatgca gtgacggcaa cggccttgaa ctgaaatgcc
    14161 cgtttacctc ccgggatttc atgaagttcc ggctcggtgg tttcgaggcc ataaagtcag
    14221 cttacatggc ccaggtgcag tacagcatgt gggtgacgcg aaaaaatgcc tggtactttg
    14281 ccaactatga cccgcgtatg aagcgtgaag gcctgcatta tgtcgtgatt gagcgggatg
    14341 aaaagtacat ggcgagtttt gacgagatcg tgccggagtt catcgaaaaa atggacgagg
    14401 cactggctga aattggtttt gtatttgggg agcaatggcg agcttaacca ggcatcaaat
    14461 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
    14521 cgctctccgc tgtcagcagc tacgctctga gcccggagag cgaccaggtg ctcggcgtgg
    14581 caagactcgc agcgaacccg
//
