LOCUS       sequence-99582-e       12162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Also referred to as mRGNndm-1. Expresses Cas9, tracrRNA and a guide 
            RNA which target the NDM-1 gene. Contains an R1162 origin and 
            chloramphenicol resistance. Can be mobilized by S17-1 lambda pir..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12162)
  AUTHORS   Citorik RJ, Mimee M, Lu TK
  TITLE     Sequence-specific antimicrobials using efficiently delivered 
            RNA-guided nucleases.
  JOURNAL   Nat Biotechnol. 2014 Sep 21. doi: 10.1038/nbt.3011.
  PUBMED    25240928
REFERENCE   2  (bases 1 to 12162)
  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 
            Biotechnol. 2014 Sep 21. doi: 10.1038/nbt.3011."
COMMENT     Sequence Label: pMM441
FEATURES             Location/Qualifiers
     source          1..12162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      11..97
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    164..295
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   296..331
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   362..397
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     rep_origin      1198..1592
                     /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            1934..2021
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             3260..4231
                     /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             4742..5581
                     /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             5568..6419
                     /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
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     terminator      complement(6633..6727)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6748..7407)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7322..7341
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7408..7510)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     7576..7594
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_RNA        complement(7680..7758)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        join(7923..7941,7942..7947,7948..7964,7965..7970,
                     7971..7996)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7923..7941
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7948..7966
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             8006..8017
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             8032..8049
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             8050..12156
                     /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             8050..12153
                     /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"
ORIGIN
        1 ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
       61 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga cctagcaccc
      121 tgcagaacta tatgattttc cgcagtatat tttagatgaa gattatttct taataactaa
      181 aaatatggta taatactctt aataaatgca gtaatacagg ggcttttcaa gactgaagtc
      241 tagctgagac aaatagtgcg attacgaaat tttttagaca aaaatagtct acgaggtttt
      301 agagctatgc tgttttgaat ggtcccaaaa cgggcagtcg cttccaacgg tttgatcgtc
      361 agttttagag ctatgctgtt ttgaatggtc ccaaaacttc agcacactga gacttgttga
      421 gttccatgtt ttagagctat gctgttttga atggtctcca ttctctagat gatgattaag
      481 gatctatttt tttgggcggg gccgcccaaa aaaatggttc gcttgctgtc ccccggggac
      541 agacgtcata ctagatatca agcgacttct cctatcccct gggaacacat caatctcacc
      601 ggagaatatc gctggccaaa gccttagcgt aggattccgc cccttcccgc aaacgacccc
      661 aaacaggaaa cgcagctgaa acgggaagct caacacccac tgacgcatgg gttgttcagg
      721 cagtacttca tcaaccagca aggcggcact ttcggccatc cgccgcgccc cacagctcgg
      781 gcagaaaccg cgacgcttac agctgaaagc gaccaggtgc tcggcgtggc aagactcgca
      841 gcgaacccgt agaaagccat gctccagccg cccgcattgg agaaattctt caaattcccg
      901 ttgcacatag cccggcaatt cctttccctg ctctgccata agcgcagcga atgccgggta
      961 atactcgtca acgatctgat agagaagggt ttgctcgggt cggtggctct ggtaacgacc
     1021 agtatcccga tcccggctgg ccgtcctggc cgccacatga ggcatgttcc gcgtccttgc
     1081 aatactgtgt ttacatacag tctatcgctt agcggaaagt tcttttaccc tcagccgaaa
     1141 tgcctgccgt tgctagacat tgccagccag tgcccgtcac tcccgtacta actgtcacga
     1201 acccctgcaa taactgtcac gcccccctgc aataactgtc acgaacccct gcaataactg
     1261 tcacgccccc aaacctgcaa acccagcagg ggcgggggct ggcggggtgt tggaaaaatc
     1321 catccatgat tatctaagaa taatccacta ggcgcggtta tcagcgccct tgtggggcgc
     1381 tgctgccctt gcccaatatg cccggccaga ggccggatag ctggtctatt cgctgcgcta
     1441 ggctacacac cgccccaccg ctgcgcggca gggggaaagg cgggcaaagc ccgctaaacc
     1501 ccacaccaaa ccccgcagaa atacgctgga gcgcttttag ccgctttagc ggcctttccc
     1561 cctacccgaa gggtgggggc gcgtgtgcag ccccgcaggg cctgtctcgg tcgatcattc
     1621 agcccggctc atccttctgg cgtggcggca gaccgaacaa ggcgcggtcg tggtcgcgtt
     1681 caaggtacgc atccattgcc gccatgagcc gatcctccgg ccactcgctg ctgttcacct
     1741 tggccaaaat catggccccc accagcacct tgcgccttgt ttcgttcttg cgctcttgct
     1801 gctgttccct tgcccgcacc cgctgaattt cggcattgat tcgcgctcgt tgttcttcga
     1861 gcttggccag ccgatccgcc gccttgttgc tccccttaac catcttgaca ccccattgtt
     1921 aatgtgctgt ctcgtaggct atcatggagg cacagcggcg gcaatcccga ccctactttg
     1981 taggggaggg cgcacttacc ggtttctctt cgagaaactg gcctaacggc cacccttcgg
     2041 gcggtgcgct ctccgagggc cattgcatgg agccgaaaag caaaagcaac agcgaggcag
     2101 catggcgatt tatcacctta cggcgaaaac cggcagcagg tcgggcggcc aatcggccag
     2161 ggccaaggcc gactacatcc agcgcgaagg caagtatgcc cgcgacatgg atgaagtctt
     2221 gcacgccgaa tccgggcaca tgccggagtt cgtcgagcgg cccgccgact actgggatgc
     2281 tgccgacctg tatgaacgcg ccaatgggcg gctgttcaag gaggtcgaat ttgccctgcc
     2341 ggtcgagctg accctcgacc agcagaaggc gctggcgtcc gagttcgccc agcacctgac
     2401 cggtgccgag cgcctgccgt atacgctggc catccatgcc ggtggcggcg agaacccgca
     2461 ctgccacctg atgatctccg agcggatcaa tgacggcatc gagcggcccg ccgctcagtg
     2521 gttcaagcgg tacaacggca agaccccgga gaagggcggg gcacagaaga ccgaagcgct
     2581 caagcccaag gcatggcttg agcagacccg cgaggcatgg gccgaccatg ccaaccgggc
     2641 attagagcgg gctggccacg acgcccgcat tgaccacaga acacttgagg cgcagggcat
     2701 cgagcgcctg cccggtgttc acctggggcc gaacgtggtg gagatggaag gccggggcat
     2761 ccgcaccgac cgggcagacg tggccctgaa catcgacacc gccaacgccc agatcatcga
     2821 cttacaggaa taccgggagg caatagacca tgaacgcaat cgacagagtg aagaaatcca
     2881 gaggcatcaa cgagttagcg gagcagatcg aaccgctggc ccagagcatg gcgacactgg
     2941 ccgacgaagc ccggcaggtc atgagccaga cccagcaggc cagcgaggcg caggcggcgg
     3001 agtggctgaa agcccagcgc cagacagggg cggcatgggt ggagctggcc aaagagttgc
     3061 gggaggtagc cgccgaggtg agcagcgccg cgcagagcgc ccggagcgcg tcgcgggggt
     3121 ggcactggaa gctatggcta accgtgatgc tggcttccat gatgcctacg gtggtgctgc
     3181 tgatcgcatc gttgctcttg ctcgacctga cgccactgac aaccgaggac ggctcgatct
     3241 ggctgcgctt ggtggcccga tgaagaacga caggactttg caggccatag gccgacagct
     3301 caaggccatg ggctgtgagc gcttcgatat cggcgtcagg gacgccacca ccggccagat
     3361 gatgaaccgg gaatggtcag ccgccgaagt gctccagaac acgccatggc tcaagcggat
     3421 gaatgcccag ggcaatgacg tgtatatcag gcccgccgag caggagcggc atggtctggt
     3481 gctggtggac gacctcagcg agtttgacct ggatgacatg aaagccgagg gccgggagcc
     3541 tgccctggta gtggaaacca gcccgaagaa ctatcaggca tgggtcaagg tggccgacgc
     3601 cgcaggcggt gaacttcggg ggcagattgc ccggacgctg gccagcgagt acgacgccga
     3661 cccggccagc gccgacagcc gccactatgg ccgcttggcg ggcttcacca accgcaagga
     3721 caagcacacc acccgcgccg gttatcagcc gtgggtgctg ctgcgtgaat ccaagggcaa
     3781 gaccgccacc gctggcccgg cgctggtgca gcaggctggc cagcagatcg agcaggccca
     3841 gcggcagcag gagaaggccc gcaggctggc cagcctcgaa ctgcccgagc ggcagcttag
     3901 ccgccaccgg cgcacggcgc tggacgagta ccgcagcgag atggccgggc tggtcaagcg
     3961 cttcggtgat gacctcagca agtgcgactt tatcgccgcg cagaagctgg ccagccgggg
     4021 ccgcagtgcc gaggaaatcg gcaaggccat ggccgaggcc agcccagcgc tggcagagcg
     4081 caagcccggc cacgaagcgg attacatcga gcgcaccgtc agcaaggtca tgggtctgcc
     4141 cagcgtccag cttgcgcggg ccgagctggc acgggcaccg gcaccccgcc agcgaggcat
     4201 ggacaggggc gggccagatt tcagcatgta gtgcttgcgt tggtactcac gcctgttata
     4261 ctatgagtac tcacgcacag aagggggttt tatggaatac gaaaaaagcg cttcagggtc
     4321 ggtctacctg atcaaaagtg acaagggcta ttggttgccc ggtggctttg gttatacgtc
     4381 aaacaaggcc gaggctggcc gcttttcagt cgctgatatg gccagcctta accttgacgg
     4441 ctgcaccttg tccttgttcc gcgaagacaa gcctttcggc cccggcaagt ttctcggtga
     4501 ctgatatgaa agaccaaaag gacaagcaga ccggcgacct gctggccagc cctgacgctg
     4561 tacgccaagc gcgatatgcc gagcgcatga aggccaaagg gatgcgtcag cgcaagttct
     4621 ggctgaccga cgacgaatac gaggcgctgc gcgagtgcct ggaagaactc agagcggcgc
     4681 agggcggggg tagtgacccc gccagcgcct aaccaccaac tgcctgcaaa ggaggcaatc
     4741 aatggctacc cataagccta tcaatattct ggaggcgttc gcagcagcgc cgccaccgct
     4801 ggactacgtt ttgcccaaca tggtggccgg tacggtcggg gcgctggtgt cgcccggtgg
     4861 tgccggtaaa tccatgctgg ccctgcaact ggccgcacag attgcaggcg ggccggatct
     4921 gctggaggtg ggcgaactgc ccaccggccc ggtgatctac ctgcccgccg aagacccgcc
     4981 caccgccatt catcaccgcc tgcacgccct tggggcgcac ctcagcgccg aggaacggca
     5041 agccgtggct gacggcctgc tgatccagcc gctgatcggc agcctgccca acatcatggc
     5101 cccggagtgg ttcgacggcc tcaagcgcgc cgccgagggc cgccgcctga tggtgctgga
     5161 cacgctgcgc cggttccaca tcgaggaaga aaacgccagc ggccccatgg cccaggtcat
     5221 cggtcgcatg gaggccatcg ccgccgatac cgggtgctct atcgtgttcc tgcaccatgc
     5281 cagcaagggc gcggccatga tgggcgcagg cgaccagcag caggccagcc ggggcagctc
     5341 ggtactggtc gataacatcc gctggcagtc ctacctgtcg agcatgacca gcgccgaggc
     5401 cgaggaatgg ggtgtggacg acgaccagcg ccggttcttc gtccgcttcg gtgtgagcaa
     5461 ggccaactat ggcgcaccgt tcgctgatcg gtggttcagg cggcatgacg gcggggtgct
     5521 caagcccgcc gtgctggaga ggcagcgcaa gagcaagggg gtgccccgtg gtgaagccta
     5581 agaacaagca cagcctcagc cacgtccggc acgacccggc gcactgtctg gcccccggcc
     5641 tgttccgtgc cctcaagcgg ggcgagcgca agcgcagcaa gctggacgtg acgtatgact
     5701 acggcgacgg caagcggatc gagttcagcg gcccggagcc gctgggcgct gatgatctgc
     5761 gcatcctgca agggctggtg gccatggctg ggcctaatgg cctagtgctt ggcccggaac
     5821 ccaagaccga aggcggacgg cagctccggc tgttcctgga acccaagtgg gaggccgtca
     5881 ccgctgaatg ccatgtggtc aaaggtagct atcgggcgct ggcaaaggaa atcggggcag
     5941 aggtcgatag tggtggggcg ctcaagcaca tacaggactg catcgagcgc ctttggaagg
     6001 tatccatcat cgcccagaat ggccgcaagc ggcaggggtt tcggctgctg tcggagtacg
     6061 ccagcgacga ggcggacggg cgcctgtacg tggccctgaa ccccttgatc gcgcaggccg
     6121 tcatgggtgg cggccagcat gtgcgcatca gcatggacga ggtgcgggcg ctggacagcg
     6181 aaaccgcccg cctgctgcac cagcggctgt gtggctggat cgaccccggc aaaaccggca
     6241 aggcttccat agataccttg tgcggctatg tctggccgtc agaggccagt ggttcgacca
     6301 tgcgcaagcg ccgccagcgg gtgcgcgagg cgttgccgga gctggtcgcg ctgggctgga
     6361 cggtaaccga gttcgcggcg ggcaagtacg acatcacccg gcccaaggcg gcaggctgac
     6421 cccccccact ctattgtaaa caagacattt ttatctttta tattcaatgg cttattttcc
     6481 tgctaattgg taataccatg aaaaatacca tgctcagaaa aggcttaaca atattttgaa
     6541 aaattgccta ctgagcgctg ccgcacagct ccataggccg ctttcctggc tttgcttcca
     6601 gatgtatgct cttctgctcc actagtgctt ggattctcac caataaaaaa cgcccggcgg
     6661 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     6721 aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg
     6781 ttgtaattca ttaagcattc tgccgacatg gaagccatca cagacggcat gatgaacctg
     6841 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     6901 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     6961 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     7021 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     7081 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     7141 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     7201 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     7261 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     7321 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     7381 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     7441 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     7501 gtgccgatca acgtctcatt ttcgccagat atcgacgtct aagaaaccat tattatcatg
     7561 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaccc tgtcgacgcg
     7621 ttttgaaagc gcttgttttt ttggtttgca gtcagagtag aatagaagta tcaaaaaaag
     7681 caccgactcg gtgccacttt ttcaagttga taacggacta gccttatttt aacttgctat
     7741 gctgttttga atggttccaa caagattatt ttataacttt tataacaaat aatcaaggag
     7801 aaattcaaag aaatttatca gccataaaac aatacttaat actatagaat gataacaaaa
     7861 taaactactt tttaaaagaa ttttgtgtta taatctattt attattaagt attgggatcg
     7921 attccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacatca
     7981 gcaggacgca ctgaccgaat tcattaaaga ggagaaaggt gcggccgcat gcatcaccat
     8041 caccatcaca tggataagaa atactcaata ggcttagata tcggcacaaa tagcgtcgga
     8101 tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat
     8161 acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag
     8221 acagcggaag cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat
     8281 cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc
     8341 tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct
     8401 atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat
     8461 ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc
     8521 ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat
     8581 aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa
     8641 gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt
     8701 aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta
     8761 tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat
     8821 ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga tttagataat
     8881 ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca
     8941 gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta
     9001 tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct
     9061 ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac
     9121 ggatatgcag gttatattga tgggggagct agccaagaag aattttataa atttatcaaa
     9181 ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat
     9241 ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt
     9301 gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt
     9361 gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt
     9421 ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat
     9481 tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac
     9541 tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat
     9601 tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca
     9661 gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga
     9721 aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt
     9781 gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg
     9841 ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag
     9901 gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa
     9961 acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact
    10021 ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa
    10081 acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc
    10141 catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc
    10201 gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt
    10261 ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa
    10321 aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg
    10381 cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa
    10441 gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa
    10501 aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat
    10561 gtcgatcaca ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta
    10621 acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa
    10681 aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca acgtaagttt
    10741 gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc
    10801 aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt
    10861 cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc
    10921 ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag
    10981 attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg
    11041 attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat
    11101 gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc
    11161 ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt
    11221 cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg
    11281 cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa
    11341 acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac
    11401 aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca
    11461 acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta
    11521 aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat
    11581 ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa
    11641 ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc
    11701 ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat
    11761 ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg
    11821 tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag tgaattttct
    11881 aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat
    11941 agagacaaac caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat
    12001 cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg
    12061 tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa
    12121 acacgcattg atttgagtca gctaggaggt gactgactcg ag
//
