LOCUS       sequence-363182-       15534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cr3 helper plasmid for recombination and Cas9 induction; harbors the
            Cas9 endonuclease and an E.coli-specific recombination system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15534)
  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 15534)
  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: cr3Ec
FEATURES             Location/Qualifiers
     source          1..15534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      377..771
                     /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            1113..1200
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2439..3410
                     /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             3921..4760
                     /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             4747..5598
                     /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"
     primer_bind     complement(5963..5980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(6097..6912)
                     /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(7184..7810)
                     /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     7715..7734
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        7826..7881
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7832..7850
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    7862..7880
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             7930..12033
                     /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             7930..12033
                     /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             12040..12072
                     /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      12090..12161
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      12216..12287
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(12302..13180)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        13207..13491
                     /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     13414..13433
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             13519..13935
                     /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             13941..14726
                     /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             14723..15403
                     /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      15418..15489
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cggcgtggca agactcgcag cgaacccgta gaaagccatg ctccagccgc ccgcattgga
       61 gaaattcttc aaattcccgt tgcacatagc ccggcaattc ctttccctgc tctgccataa
      121 gcgcagcgaa tgccgggtaa tactcgtcaa cgatctgata gagaagggtt tgctcgggtc
      181 ggtggctctg gtaacgacca gtatcccgat cccggctggc cgtcctggcc gccacatgag
      241 gcatgttccg cgtccttgca atactgtgtt tacatacagt ctatcgctta gcggaaagtt
      301 cttttaccct cagccgaaat gcctgccgtt gctagacatt gccagccagt gcccgtcact
      361 cccgtactaa ctgtcacgaa cccctgcaat aactgtcacg cccccctgca ataactgtca
      421 cgaacccctg caataactgt cacgccccca aacctgcaaa cccagcaggg gcgggggctg
      481 gcggggtgtt ggaaaaatcc atccatgatt atctaagaat aatccactag gcgcggttat
      541 cagcgccctt gtggggcgct gctgcccttg cccaatatgc ccggccagag gccggatagc
      601 tggtctattc gctgcgctag gctacacacc gccccaccgc tgcgcggcag ggggaaaggc
      661 gggcaaagcc cgctaaaccc cacaccaaac cccgcagaaa tacgctggag cgcttttagc
      721 cgctttagcg gcctttcccc ctacccgaag ggtgggggcg cgtgtgcagc cccgcagggc
      781 ctgtctcggt cgatcattca gcccggctca tccttctggc gtggcggcag accgaacaag
      841 gcgcggtcgt ggtcgcgttc aaggtacgca tccattgccg ccatgagccg atcctccggc
      901 cactcgctgc tgttcacctt ggccaaaatc atggccccca ccagcacctt gcgccttgtt
      961 tcgttcttgc gctcttgctg ctgttccctt gcccgcaccc gctgaatttc ggcattgatt
     1021 cgcgctcgtt gttcttcgag cttggccagc cgatccgccg ccttgttgct ccccttaacc
     1081 atcttgacac cccattgtta atgtgctgtc tcgtaggcta tcatggaggc acagcggcgg
     1141 caatcccgac cctactttgt aggggagggc gcacttaccg gtttctcttc gagaaactgg
     1201 cctaacggcc acccttcggg cggtgcgctc tccgagggcc attgcatgga gccgaaaagc
     1261 aaaagcaaca gcgaggcagc atggcgattt atcaccttac ggcgaaaacc ggcagcaggt
     1321 cgggcggcca atcggccagg gccaaggccg actacatcca gcgcgaaggc aagtatgccc
     1381 gcgacatgga tgaagtcttg cacgccgaat ccgggcacat gccggagttc gtcgagcggc
     1441 ccgccgacta ctgggatgct gccgacctgt atgaacgcgc caatgggcgg ctgttcaagg
     1501 aggtcgaatt tgccctgccg gtcgagctga ccctcgacca gcagaaggcg ctggcgtccg
     1561 agttcgccca gcacctgacc ggtgccgagc gcctgccgta tacgctggcc atccatgccg
     1621 gtggcggcga gaacccgcac tgccacctga tgatcncyga gcggatcaat gacggcatcg
     1681 agcggcccgc cgctcagtgg ttcaagcggt acaacggcaa gaccccggag aagggcgggg
     1741 cacaaaagac cgaagcgctc aagcccaagg catggcttga gcagacccgc gaggcatggg
     1801 ccgaccatgc caaccgggca ttagagcggg ctggccacga cgcccgcatt gaccacagaa
     1861 cacttgaggc gcagggcatc gagcgcctgc ccggtgttca cctggggccg aacgtggtgg
     1921 agatggaagg ccggggcatc cgcaccgacc gggcagacgt ggccctgaac atcgacaccg
     1981 ccaacgccca gatcatcgac ttacaggaat accgggaggc aatagaccat gaacgcaatc
     2041 gacagagtga agaaatccag aggcatcaac gagttagcgg agcagatcga accgctggcc
     2101 cagagcatgg cgacactggc cgacgaagcc cggcaggtca tgagccagac ccagcaggcc
     2161 agcgaggcgc aggcggcgga gtggctgaaa gcccagcgcc agacaggggc ggcatgggtg
     2221 gagctggcca aagagttgcg ggaggtagcc gccgaggtga gcagcgccgc gcagagcgcc
     2281 cggagcgcgt cgcgggggtg gcactggaag ctatggctaa ccgtgatgct ggcttccatg
     2341 atgcctacgg tggtgctgct gatcgcatcg ttgctcttgc tcgacctgac gccactgaca
     2401 accgaggacg gctcgatctg gctgcgcttg gtggcccgat gaagaacgac aggactttgc
     2461 aggccatagg ccgacagctc aaggccatgg gctgtgagcg cttcgatatc ggcgtcaggg
     2521 acgccaccac cggccagatg atgaaccggg aatggtcagc cgccgaagtg ctccagaaca
     2581 cgccatggct caagcggatg aatgcccagg gcaatgacgt gtatatcagg cccgccgagc
     2641 aggagcggca tggtctggtg ctggtggacg acctcagcga gtttgacctg gatgacatga
     2701 aagccgaggg ccgggagcct gccctggtag tggaaaccag cccgaagaac tatcaggcat
     2761 gggtcaaggt ggccgacgcc gcaggcggtg aacttcgggg gcagattgcc cggacgctgg
     2821 ccagcgagta cgacgccgac ccggccagcg ccgacagccg ccactatggc cgcttggcgg
     2881 gcttcaccaa ccgcaaggac aagcacacca cccgcgccgg ttatcagccg tgggtgctgc
     2941 tgcgtgaatc caagggcaag accgccaccg ctggcccggc gctggtgcag caggctggcc
     3001 agcagatcga gcaggcccag cggcagcagg agaaggcccg caggctggcc agcctcgaac
     3061 tgcccgagcg gcagcttagc cgccaccggc gcacggcgct ggacgagtac cgcagcgaga
     3121 tggccgggct ggtcaagcgc ttcggtgatg acctcagcaa gtgcgacttt atcgccgcgc
     3181 agaagctggc cagccggggc cgcagtgccg aggaaatcgg caaggccatg gccgaggcca
     3241 gcccagcgct ggcagagcgc aagcccggcc acgaagcgga ttacatcgag cgcaccgtca
     3301 gcaaggtcat gggtctgccc agcgtccagc ttgcgcgggc cgagctggca cgggcaccgg
     3361 caccccgcca gcgaggcatg gacaggggcg ggccagattt cagcatgtag tgcttgcgtt
     3421 ggtactcacg cctgttatac tatgagtact cacgcacaga agggggtttt atggaatacg
     3481 aaaaaagcgc ttcagggtcg gtctacctga tcaaaagtga caagggctat tggttgcccg
     3541 gtggctttgg ttatacgtca aacaaggccg aggctggccg cttttcagtc gctgatatgg
     3601 ccagccttaa ccttgacggc tgcaccttgt ccttgttccg cgaagataag cctttcggcc
     3661 ccggcaagtt tctcggtgac tgatatgaaa gaccaaaagg acaagcagac cggcgacctg
     3721 ctggccagcc ctgacgctgt acgccaagcg cgatatgccg agcgcatgaa ggccaaaggg
     3781 atgcgtcagc gcaagttctg gctgaccgac gacgaatacg aggcgctgcg cgagtgcctg
     3841 gaagaactca gagcggcgca gggcgggggt agtgaccccg ccagcgccta accaccaact
     3901 gcctgcaaag gaggcaatca atggctaccc ataagcctat caatattctg gaggcgttcg
     3961 cagcagcgcc gccaccgctg gactacgttt tgcccaacat ggtggccggt acggtcgggg
     4021 cgctggtgtc gcccggtggt gccggtaaat ccatgctggc cctgcaactg gccgcacaga
     4081 ttgcaggcgg gccggatctg ctggaggtgg gcgaactgcc caccggcccg gtgatctacc
     4141 tgcccgccga agatccgccc accgccattc atcaccgcct gcacgccctt ggggcgcacc
     4201 tcagcgccga ggaacggcaa gccgtggctg acggcctgct gatccagccg ctgatcggca
     4261 gcctgcccaa catcatggcc ccggagtggt tcgacggcct caagcgcgcc gccgagggcc
     4321 gccgcctgat ggtgctggac acgctgcgcc ggttccacat cgaggaagaa aacgccagcg
     4381 gccccatggc ccaggtcatc ggtcgcatgg aggccatcgc cgccgatacc gggtgctcta
     4441 tcgtgttcct gcaccatgcc agcaagggcg cggccatgat gggcgcaggc gaccagcagc
     4501 aggccagccg gggcagctcg gtactggtcg ataacatccg ctggcagtcc tacctgtcga
     4561 gcatgaccag cgccgaggcc gaggaatggg gtgtggacga cgaccagcgc cggttcttcg
     4621 tccgcttcgg tgtgagcaag gccaactatg gcgcaccgtt cgctgatcgg tggttcaggc
     4681 ggcatgacgg cggggtgctc aagcccgccg tgctggagag gcagcgcaag agcaaggggg
     4741 tgccccgtgg tgaagcctaa gaacaagcac agcctcagcc acgtccggca cgacccggcg
     4801 cactgtctgg cccccggcct gttccgtgcc ctcaagcggg gcgagcgcaa gcgcagcaag
     4861 ctggacgtga cgtatgacta cggcgacggc aagcggatcg agttcagcgg cccggagccg
     4921 ctgggcgctg atgatctgcg catcctgcaa gggctggtgg ccatggctgg gcctaatggc
     4981 ctagtgcttg gcccggaacc caagaccgaa ggcggacggc agctccggct gttcctggaa
     5041 cccaagtggg aggccgtcac cgctgatgcc atggtggtca aaggtagcta tcgggcgctg
     5101 gcaaaggaaa tcggggcaga ggtcgatagt ggtggggcgc tcaagcacat acaggactgc
     5161 atcgagcgcc tttggaaggt atccatcatc gcccagaatg gccgcaagcg gcaggggttt
     5221 cggctgctgt cggagtacgc cagcgacgag gcggacgggc gcctgtacgt ggccctgaac
     5281 cccttgatcg cgcaggccgt catgggtggc ggccagcatg tgcgcatcag catggacgag
     5341 gtgcgggcgc tggacagcga aaccgcccgc ctgctgcacc agcggctgtg tggctggatc
     5401 gaccccggca aaaccggcaa ggcttccata gataccttgt gcggctatgt ctggccgtca
     5461 gaggccagtg gttcgaccat gcgcaagcgc cgccagcggg tgcgcgaggc gttgccggag
     5521 ctggtcgcgc tgggctggac ggtaaccgag ttcgcggcgg gcaagtacga catcacccgg
     5581 cccaaggcgg caggctgacc ccccccactc tattgtaaac aagacatttt tatcttttat
     5641 attcaatggc ttattttcct gctaattggt aataccatga aaaataccat gctcagaaaa
     5701 ggcttaacaa tattttgaaa aattgcctac tgagcgctgc cgcacagctc cataggccgc
     5761 tttcctggct ttgcttccag atgtatgctc ttctgctccc gaacgccagc aagacgtagc
     5821 ccagcgcgtc ggccagcttg caattcgcgc taacttacat tcgctagccg cagctacgct
     5881 ctgagtgcag cgcgggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
     5941 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6001 ggcgagcggt atcagctcac tcaaaggcgg taatctcgag tcccgtcaag tcagcgtaat
     6061 gctctgccag tgttacaacc aattaaccaa ttctgattaa aagaactcgt ccagcatcag
     6121 gtgaaattgc agtttgttca tgtctgggtt atcgatgccg tatttttgaa acagacgttt
     6181 ttgcagagat gggctaaatt cacccagaca gttccacaga attgccagat cttggtaacg
     6241 atctgcaata ccaacacgac caacatcaat acaaccgatc agtttaccct cgtcaaaaat
     6301 caggttatcc agagaaaaat caccgtgggt aacaacgcta tctggagaaa atggcagcag
     6361 tttgtgcatt tctttccaaa cttgttcaac tggccaaccg ttacgttcat catcaaaatc
     6421 agatgcatca accagaccgt tgttcatacg agattgtgct tgtgccagac gaaaaacacg
     6481 atcgctgtta aatggacagt tacaaactgg aatagagtgc agacgacgca gaaaaactgc
     6541 cagtgcatca acaatgtttt caccagaatc tgggtattct tccagaactt gaaatgcggt
     6601 tttacctgga attgcggtgg tcagcagcca tgcatcatct ggggtacgaa taaagtgttt
     6661 aatggttggc agtggcataa attcggtcag ccagtttaaa cgaaccattt catcggtaac
     6721 atcgtttgca acagaacctt taccgtgttt cagaaacagt tctggtgcat ctggtttacc
     6781 gtaaagacgg taaatggttg caccagattg accaacgtta tcacgtgccc atttgtaacc
     6841 gtacagatct gcatccatgt tagagttcag acgtggacga gaacaagagg tttcacgttg
     6901 aatgtgagac ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6961 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     7021 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttaacctccg
     7081 ccttcagagc gtagctgcgc caggagtttt gttatcaata aaaaaggccc cccgttaggg
     7141 aggcttattg ttcgtcccat cacggaaaaa ggttatgctg cttttaagac ccactttcac
     7201 atttaagttg tttttctaat ccgcatatga tcaattcaag gccgaataag aaggctggct
     7261 ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc atactatcag
     7321 tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc aatacgcaac
     7381 ctaaagtaaa atgccccaca gcgctgagtg catataatgc attctctagt gaaaaacctt
     7441 gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct gtaggccgtg
     7501 tacctaaatg tacttttgct ccatcgcgat gacttagtaa agcacatcta aaacttttag
     7561 cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag tgagtatggt
     7621 gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt tttacatgcc
     7681 aatacaatgt aggctgctct acacctagcc tctgggcgag tttacgggtt gttaaacctt
     7741 cgattccgac ctcattaagc agctctaatg cgctgctaat cactctactt ttatctaatc
     7801 gagacatcat taattcctaa tttttgttga cactctatca ttgatagagt tattttacca
     7861 ctccctatca gtgatagaga aaagtgaata ctaaagaatt caaaagatct aaagaggact
     7921 tcggatctaa tggataagaa atactcaata ggcttagata tcggcacaaa tagcgtcgga
     7981 tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat
     8041 acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag
     8101 acagcggaag cgactcgtct gaaacggaca gctcgtagaa ggtatacacg tcggaagaat
     8161 cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc
     8221 tttcatcgac ttgaagagtc ttttttggtg gaagaagata agaagcatga acgtcatcct
     8281 atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat
     8341 ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc
     8401 ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat
     8461 aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa
     8521 gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt
     8581 aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta
     8641 tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat
     8701 ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga tttagataat
     8761 ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca
     8821 gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta
     8881 tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct
     8941 ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac
     9001 ggatatgcag gttatattga tgggggagct agccaagaag aattttataa atttatcaaa
     9061 ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat
     9121 ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt
     9181 gagctgcatg ctattttgag aaggcaagaa gatttttatc catttttaaa agacaatcgt
     9241 gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt
     9301 ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat
     9361 tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac
     9421 tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat
     9481 tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca
     9541 gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga
     9601 aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt
     9661 gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg
     9721 ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag
     9781 gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa
     9841 acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact
     9901 ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa
     9961 acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc
    10021 catgatgata gtttgacatt taaagaagat attcaaaaag cacaagtgtc tggacaaggc
    10081 gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt
    10141 ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa
    10201 aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg
    10261 cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa
    10321 gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa
    10381 aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat
    10441 gtcgatcaca ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta
    10501 acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa
    10561 aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca acgtaagttt
    10621 gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc
    10681 aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt
    10741 cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc
    10801 ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag
    10861 attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg
    10921 attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat
    10981 gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc
    11041 ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt
    11101 cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg
    11161 cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa
    11221 acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac
    11281 aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca
    11341 acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta
    11401 aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat
    11461 ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa
    11521 ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc
    11581 ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat
    11641 ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg
    11701 tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag tgaattttct
    11761 aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat
    11821 agagacaaac caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat
    11881 cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg
    11941 tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa
    12001 acacgcattg atttgagtca gctaggaggt gacgctttag ctgctaacga cgaaaactac
    12061 gctctggctg cttaggggta accaggcatc aaataaaacg aaaggctcag tcgaaagact
    12121 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ccgctggacg cagctacgct
    12181 ctgagtacgg agctcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga
    12241 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcgag
    12301 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
    12361 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
    12421 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
    12481 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
    12541 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
    12601 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
    12661 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
    12721 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
    12781 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
    12841 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
    12901 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
    12961 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
    13021 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
    13081 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
    13141 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
    13201 tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact
    13261 ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg
    13321 ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc
    13381 acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat
    13441 tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca tatactagag
    13501 aaagagggga aataatcaat ggatattaat actgaaactg agatcaagca aaagcattca
    13561 ctaaccccct ttcctgtttt cctaatcagc ccggcatttc gcgggcgata ttttcacagc
    13621 tatttcagga gttcagccat gaacgcttat tacattcagg atcgtcttga ggctcagagc
    13681 tgggcgcgtc actaccagca gctcgcccgt gaagagaaag aggcagaact ggcagacgac
    13741 atggaaaaag gcctgcccca gcacctgttt gaatcgctat gcatcgatca tttgcaacgc
    13801 cacggggcca gcaaaaaatc cattacccgt gcgtttgatg acgatgttga gtttcaggag
    13861 cgcatggcag aacacatccg gtacatggtt gaaaccattg ctcaccacca ggttgatatt
    13921 gattcagagg tataaaacga atgagtactg cactcgcaac gctggctggg aagctggctg
    13981 aacgtgtcgg catggattct gtcgacccac aggaactgat caccactctt cgccagacgg
    14041 catttaaagg tgatgccagc gatgcgcagt tcatcgcatt actgatcgtt gccaaccagt
    14101 acggccttaa tccgtggacg aaagaaattt acgcctttcc tgataagcag aatggcatcg
    14161 ttccggtggt gggcgttgat ggctggtccc gcatcatcaa tgaaaaccag cagtttgatg
    14221 gcatggactt tgagcaggac aatgaatcct gtacatgccg gatttaccgc aaggaccgta
    14281 atcatccgat ctgcgttacc gaatggatgg atgaatgccg ccgcgaacca ttcaaaactc
    14341 gcgaaggcag agaaatcacg gggccgtggc agtcgcatcc caaacggatg ttacgtcata
    14401 aagccatgat tcagtgtgcc cgtctggcct tcggatttgc tggtatctat gacaaggatg
    14461 aagccgagcg cattgtcgaa aatactgcat acactgcaga acgtcagccg gaacgcgaca
    14521 tcactccggt taacgatgaa accatgcagg agattaacac tctgctgatc gccctggata
    14581 aaacatggga tgacgactta ttgccgctct gttcccagat atttcgccgc gacattcgtg
    14641 catcgtcaga actgacacag gccgaagcag taaaagctct tggattcctg aaacagaaag
    14701 ccgcagagca gaaggtggca gcatgacacc ggacattatc ctgcagcgta ccgggatcga
    14761 tgtgagagct gtcgaacagg gggatgatgc gtggcacaaa ttacggctcg gcgtcatcac
    14821 cgcttcagaa gttcacaacg tgatagcaaa accccgctcc ggaaagaagt ggcctgacat
    14881 gaaaatgtcc tacttccaca ccctgcttgc tgaggtttgc accggtgtgg ctccggaagt
    14941 taacgctaaa gcactggcct ggggaaaaca gtacgagaac gacgccagaa ccctgtttga
    15001 attcacttcc ggcgtgaatg ttactgaatc cccgatcatc tatcgcgacg aaagtatgcg
    15061 taccgcctgc tctcccgatg gtttatgcag tgacggcaac ggccttgaac tgaaatgccc
    15121 gtttacctcc cgggatttca tgaagttccg gctcggtggt ttcgaggcca taaagtcagc
    15181 ttacatggcc caggtgcagt acagcatgtg ggtgacgcga aaaaatgcct ggtactttgc
    15241 caactatgac ccgcgtatga agcgtgaagg cctgcattat gtcgtgattg agcgggatga
    15301 aaagtacatg gcgagttttg acgagatcgt gccggagttc atcgaaaaaa tggacgaggc
    15361 actggctgaa attggttttg tatttgggga gcaatggcga tgagcttaac caggcatcaa
    15421 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
    15481 aacgctctcc gctgtcagca gctacgctct gagcccggag agcgaccagg tgct
//
