LOCUS       sequence-434126-       12066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombineering plasmid with constitutively expressed cas9 and the 
            araBAD promoter expressing alpha-beta-gamma on a BBR1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12066)
  AUTHORS   Venkataraman M, Ynigez-Gutierrez A, Infante V, MacIntyre A, 
            Fernandes-Junior PI, Ane JM, Pfleger B
  TITLE     Synthetic Biology Toolbox for Nitrogen-Fixing Soil Microbes.
  JOURNAL   ACS Synth Biol. 2023 Dec 15;12(12):3623-3634. doi: 
            10.1021/acssynbio.3c00414. Epub 2023 Nov 21.
  PUBMED    37988619
REFERENCE   2  (bases 1 to 12066)
  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"; doi: 
            "10.1021/acssynbio.3c00414"; journalName: "ACS Synth Biol"; date: 
            "2023-12-15- 15"; volume: "12"; issue: "12"; pages: "3623-3634"
COMMENT     Sequence Label: pMVV205
FEATURES             Location/Qualifiers
     source          1..12066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(122..784)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      785..1556
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(1590..1684)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1944..2477)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEVVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2666..2671,2672..2688,2689..2694))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(2909..2932)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2942..7045)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(2942..7045)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     promoter        complement(7081..7115)
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_feature    7116..7137
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(7187..7233)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7386..7403
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(7459..7476)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(7492..7521)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_RNA        complement(7542..7617)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    7648..7666
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7673..7691
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7748..7825
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7826..8449
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(7899..7918)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      8583..8614
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(8665..9543)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        9570..9854
                     /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     9777..9796
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             9881..9889
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             9898..10314
                     /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             10320..11105
                     /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             11102..11782
                     /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      11783..12027
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
ORIGIN
        1 ttatacctag ggcgatgcag gtggctgctg aacccccagc cggaactgac cccacaaggc
       61 ccctaggcta cagccgatag tctggaacag cgcacttacg ggttgctgcg caacccaagt
      121 gctaccggcg cggcagcgtg acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa
      181 aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt
      241 cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg ggcggctcct
      301 cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt
      361 cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga
      421 acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca ttgaccacgt
      481 aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc tcggccacca
      541 agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg gaaagtgtct
      601 tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga tgcagcagca
      661 ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg
      721 tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg
      781 ccatgcttat ctccatgcgg taggggtgcc gcacggttgc ggcaccatgc gcaatcagct
      841 gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag tcaattacag
      901 attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc tgttgcgtac
      961 cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata tctagttctt
     1021 tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc ggctccccct
     1081 ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggcctta ggccttgccc
     1141 aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg gggggcaggc
     1201 gggcgggctt cgcccttcga ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt
     1261 caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc
     1321 aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag aaaattaaaa
     1381 aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg tggtcgaagg
     1441 ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc
     1501 agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgctcgc
     1561 ggccatcgtc cacatatcct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     1621 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     1681 agtcaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt gaatcgggat
     1741 atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct gacggaacag
     1801 cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg cgcacatttc
     1861 cccgaaaagt gccacctgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     1921 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     1981 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     2041 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     2101 agcgcggtgg caatgccctg cctccggtgc tctccggaga ctgcgagatc atagatatag
     2161 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     2221 acttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     2281 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     2341 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     2401 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     2461 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     2521 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
     2581 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     2641 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt
     2701 tcgtgccttc atccgtttcc acggtgtgcg tccatgggca aatattatac gcaaggcgac
     2761 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2821 ggcagaatgc ttaatgaatt acaacagttt ttatgcatgc gcagtccagt tacgctggag
     2881 tctgaggctc gtcctgaatg atatctcatt tatcatcatc atctttataa tcaccgctgc
     2941 cgtcacctcc tagctgactc aaatcaatgc gtgtttcata aagaccagtg atggattgat
     3001 ggataagagt ggcatctaaa acttcttttg tagacgtata tcgtttacga tcaattgttg
     3061 tatcaaaata tttaaaagca gcgggagctc caagattcgt caacgtaaat aaatgaataa
     3121 tattttctgc ttgttcacgt attggtttgt ctctatgttt gttatatgca ctaagaactt
     3181 tatctaaatt ggcatctgct aaaataacac gcttagaaaa ttcactgatt tgctcaataa
     3241 tctcatctaa ataatgctta tgctgctcca caaacaattg tttttgttcg ttatcttctg
     3301 gactaccctt caacttttca taatgactag ctaaatataa aaaattcaca tatttgcttg
     3361 gcagagccag ctcatttcct ttttgtaatt ctccggcact agccagcatc cgtttacgac
     3421 cgttttctaa ctcaaaaaga ctatatttag gtagtttaat gattaagtct tttttaactt
     3481 ccttatatcc tttagcttct aaaaagtcaa tcggattttt ttcaaaggaa cttctttcca
     3541 taattgtgat ccctagtaac tctttaacgg attttaactt cttcgatttc cctttttcca
     3601 ccttagcaac cactaggact gaataagcta ccgttggact atcaaaacca ccatattttt
     3661 ttggatccca gtctttttta cgagcaataa gcttgtccga atttcttttt ggtaaaattg
     3721 actccttgga gaatccgcct gtctgtactt ctgttttctt gacaatattg acttggggca
     3781 tggacaatac tttgcgcact gtggcaaaat ctcgcccttt atcccagaca atttctccag
     3841 tttccccatt agtttcgatt agagggcgtt tgcgaatctc tccatttgca agtgtaattt
     3901 ctgttttgaa gaagttcatg atattagagt aaaagaaata ttttgcggtt gctttgccta
     3961 tttcttgctc agacttagca atcattttac gaacatcata aactttataa tcaccataga
     4021 caaactccga ttcaagtttt ggatatttct taatcaaagc agttccaacg acggcattta
     4081 gatacgcatc atgggcatga tggtaattgt taatctcacg tactttatag aattggaaat
     4141 cttttcggaa gtcagaaact aatttagatt ttaaggtaat cactttaacc tctcgaataa
     4201 gtttatcatt ttcatcgtat ttagtattca tgcgactatc caaaatttgt gccacatgct
     4261 tagtgatttg gcgagtttca accaattggc gtttgataaa accagcttta tcaagttcac
     4321 tcaaacctcc acgttcagct ttcgttaaat tatcaaactt acgttgagtg attaacttgg
     4381 cgtttagaag ttgtctccaa tagtttttca tctttttgac tacttcttca cttggaacgt
     4441 tatccgattt accacgattt ttatcagaac gcgttaagac cttattgtct attgaatcgt
     4501 ctttaaggaa actttgtgga acaatgtgat cgacatcata atcacttaaa cgattaatat
     4561 ctaattcttg gtccacatac atgtctcttc cattttggag ataatagaga tagagctttt
     4621 cattttgcaa ttgagtattt tcaacaggat gctctttaag aatctgactt cctaattctt
     4681 tgataccttc ttcgattcgt ttcatacgct ctcgcgaatt tttctggccc ttttgagttg
     4741 tctgattttc acgtgccatt tcaataacga tattttctgg cttatgccgc cccattactt
     4801 tgaccaattc atcaacaact tttacagtct gtaaaatacc ttttttaata gcagggctac
     4861 cagctaaatt tgcaatatgt tcatgtaaac tatcgccttg tccagacact tgtgcttttt
     4921 gaatgtcttc tttaaatgtc aaactatcat catggatcag ctgcataaaa ttgcgattgg
     4981 caaaaccatc tgatttcaaa aaatctaata ttgttttgcc agattgctta tccctaatac
     5041 cattaatcaa ttttcgagac aaacgtcccc aaccagtata acggcgacgt ttaagctgtt
     5101 tcatcacctt atcatcaaag aggtgagcat atgttttaag tctttcctca atcatctccc
     5161 tatcttcaaa taaggtcaat gttaaaacaa tatcctctaa gatatcttca ttttcttcat
     5221 tatccaaaaa atctttatct ttaataattt ttagcaaatc atggtaggta cctaatgaag
     5281 cattaaatct atcttcaact cctgaaattt caacactatc aaaacattct atttttttga
     5341 aataatcttc ttttaattgc ttaacggtta cttttcgatt tgttttgaag agtaaatcaa
     5401 caatggcttt cttctgttca cctgaaagaa atgctggttt tcgcattcct tcagtaacat
     5461 atttgacctt tgtcaattcg ttataaaccg taaaatactc ataaagcaaa ctatgttttg
     5521 gtagtacttt ttcatttgga agatttttat caaagtttgt catgcgttca ataaatgatt
     5581 gagctgaagc acctttatcg acaacttctt caaaattcca tggggtaatt gtttcttcag
     5641 acttccgagt catccatgca aaacgactat tgccacgcgc caatggacca acataataag
     5701 gaattcgaaa agtcaagatt ttttcaatct tctcacgatt gtcttttaaa aatggataaa
     5761 agtcttcttg tcttctcaaa atagcatgca gctcacccaa gtgaatttga tggggaatag
     5821 agccgttgtc aaaggtccgt tgcttgcgca gcaaatcttc acgatttagt ttcaccaata
     5881 attcctcagt accatccatt ttttctaaaa ttggtttgat aaatttataa aattcttctt
     5941 ggctagctcc cccatcaata taacctgcat atccgttttt tgattgatca aaaaagattt
     6001 ctttatactt ttctggaagt tgttgtcgaa ctaaagcttt taaaagagtc aagtcttgat
     6061 gatgttcatc gtagcgttta atcattgaag ctgatagggg agccttagtt atttcagtat
     6121 ttactcttag gatatctgaa agtaaaatag catctgataa attcttagct gccaaaaaca
     6181 aatcagcata ttgatctcca atttgcgcca ataaattatc taaatcatca tcgtaagtat
     6241 cttttgaaag ctgtaattta gcatcttctg ccaaatcaaa atttgattta aaattagggg
     6301 tcaaacccaa tgacaaagca atgagattcc caaataagcc atttttcttc tcaccgggga
     6361 gctgagcaat gagattttct aatcgtcttg atttactcaa tcgtgcagaa agaatcgctt
     6421 tagcatctac tccacttgcg ttaatagggt tttcttcaaa taattgattg taggtttgta
     6481 ccaactggat aaatagtttg tccacatcac tattatcagg atttaaatct ccctcaatca
     6541 aaaaatgacc acgaaactta atcatatgcg ctaaggccaa atagattaag cgcaaatccg
     6601 ctttatcagt agaatctacc aatttttttc gcagatgata gatagttgga tatttctcat
     6661 gataagcaac ttcatctact atatttccaa aaataggatg acgttcatgc ttcttgtctt
     6721 cttccaccaa aaaagactct tcaagtcgat gaaagaaact atcatctact ttcgccatct
     6781 catttgaaaa aatctcctgt agataacaaa tacgattctt ccgacgtgta taccttctac
     6841 gagctgtccg tttgagacga gtcgcttccg ctgtctctcc actgtcaaat aaaagagccc
     6901 ctataagatt ttttttgata ctgtggcggt ctgtatttcc cagaaccttg aactttttag
     6961 acggaacctt atattcatca gtgatcaccg cccatccgac gctatttgtg ccgatatcta
     7021 agcctattga gtatttctta tccattgatt gtacctcctg gtcagtgcgt cctgctgata
     7081 gctagcatag tacctaggac tgagctagcc gtaaactcta gaagcggccg cgaattcaac
     7141 tgcagtctag actcgagtaa ggatccagtt caccgacaaa caacagataa aacgaaaggc
     7201 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
     7261 ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt
     7321 caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc
     7381 gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cagccaagct
     7441 ggagaccgtt taaactcaat gatgatgatg atgatggtcg acggcgctat tcagatcctc
     7501 ttctgagatg agtttttgtt cgggcccaag cttcaaaaaa agcaccgact cggtgccact
     7561 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaacgga
     7621 agctccctcg tgcgctcgtg ctcagtatct ctatcactga tagggatgtc aatctctatc
     7681 actgataggg actcgagcta tgctactccg tcaagccgtc aattgtctga ttcgttacca
     7741 actcgaggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
     7801 agagagtcaa ttcagggtgg tgaatatgtc tagattagat aaaagtaaag tgattaacag
     7861 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
     7921 gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct
     7981 cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg
     8041 ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag
     8101 tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac
     8161 tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata
     8221 tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca
     8281 agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca
     8341 agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt
     8401 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaat accaccgtca
     8461 aaaaaaacgg cgctttttag cgccgttttt atttttcaac cttaagcttg aattcgttga
     8521 cgaattctct agatatcgct caatactgac catttaaatc atacctgacc tccatagcag
     8581 aaagtcaaaa acctccgacc ggaggctttt gacttgaggg ggatccggca catttccccg
     8641 aaaagtgcca cctgcatcga tttattatga caacttgacg gctacatcat tcactttttc
     8701 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
     8761 gaagtagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
     8821 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
     8881 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
     8941 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
     9001 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
     9061 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
     9121 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
     9181 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
     9241 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
     9301 gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg
     9361 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
     9421 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
     9481 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
     9541 catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt gcatcagaca
     9601 ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa ccccgcttat
     9661 taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt
     9721 ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac tttgctatgc
     9781 catagcattt ttatccataa gattagcgga tcctacctga cgctttttat cgcaactctc
     9841 tactgtttct ccatacccgt ttttttggga attcgagctc taaggaggtt ataaaaaatg
     9901 gatattaata ctgaaactga gatcaagcaa aagcattcac taaccccctt tcctgttttc
     9961 ctaatcagcc cggcatttcg cgggcgatat tttcacagct atttcaggag ttcagccatg
    10021 aacgcttatt acattcagga tcgtcttgag gctcagagct gggcgcgtca ctaccagcag
    10081 ctcgcccgtg aagagaaaga ggcagaactg gcagacgaca tggaaaaagg cctgccccag
    10141 cacctgtttg aatcgctatg catcgatcat ttgcaacgcc acggggccag caaaaaatcc
    10201 attacccgtg cgtttgatga cgatgttgag tttcaggagc gcatggcaga acacatccgg
    10261 tacatggttg aaaccattgc tcaccaccag gttgatattg attcagaggt ataaaacgaa
    10321 tgagtactgc actcgcaacg ctggctggga agctggctga acgtgtcggc atggattctg
    10381 tcgacccaca ggaactgatc accactcttc gccagacggc atttaaaggt gatgccagcg
    10441 atgcgcagtt catcgcatta ctgatcgttg ccaaccagta cggccttaat ccgtggacga
    10501 aagaaattta cgcctttcct gataagcaga atggcatcgt tccggtggtg ggcgttgatg
    10561 gctggtcccg catcatcaat gaaaaccagc agtttgatgg catggacttt gagcaggaca
    10621 atgaatcctg tacatgccgg atttaccgca aggaccgtaa tcatccgatc tgcgttaccg
    10681 aatggatgga tgaatgccgc cgcgaaccat tcaaaactcg cgaaggcaga gaaatcacgg
    10741 ggccgtggca gtcgcatccc aaacggatgt tacgtcataa agccatgatt cagtgtgccc
    10801 gtctggcctt cggatttgct ggtatctatg acaaggatga agccgagcgc attgtcgaaa
    10861 atactgcata cactgcagaa cgtcagccgg aacgcgacat cactccggtt aacgatgaaa
    10921 ccatgcagga gattaacact ctgctgatcg ccctggataa aacatgggat gacgacttat
    10981 tgccgctctg ttcccagata tttcgccgcg acattcgtgc atcgtcagaa ctgacacagg
    11041 ccgaagcagt aaaagctctt ggattcctga aacagaaagc cgcagagcag aaggtggcag
    11101 catgacaccg gacattatcc tgcagcgtac cgggatcgat gtgagagctg tcgaacaggg
    11161 ggatgatgcg tggcacaaat tacggctcgg cgtcatcacc gcttcagaag ttcacaacgt
    11221 gatagcaaaa ccccgctccg gaaagaagtg gcctgacatg aaaatgtcct acttccacac
    11281 cctgcttgct gaggtttgca ccggtgtggc tccggaagtt aacgctaaag cactggcctg
    11341 gggaaaacag tacgagaacg acgccagaac cctgtttgaa ttcacttccg gcgtgaatgt
    11401 tactgaatcc ccgatcatct atcgcgacga aagtatgcgt accgcctgct ctcccgatgg
    11461 tttatgcagt gacggcaacg gccttgaact gaaatgcccg tttacctccc gggatttcat
    11521 gaagttccgg ctcggtggtt tcgaggccat aaagtcagct tacatggccc aggtgcagta
    11581 cagcatgtgg gtgacgcgaa aaaatgcctg gtactttgcc aactatgacc cgcgtatgaa
    11641 gcgtgaaggc ctgcattatg tcgtgattga gcgggatgaa aagtacatgg cgagttttga
    11701 cgagatcgtg ccggagttca tcgaaaaaat ggacgaggca ctggctgaaa ttggttttgt
    11761 atttggggag caatggcgat gacgcatcct cacgataata tccgggtagg cgcaatcact
    11821 ttcgtctact ccgttacaaa gcgaggctgg gtatttcccg gcctttctgt tatccgaaat
    11881 ccactgaaag cacagcggct ggctgaggag ataaataata aacgaggggc tgtatgcaca
    11941 aagcatcttc tgttgagtta agaacgagta tcgagatggc acatagcctt gctcaaattg
    12001 gaatcaggtt tgtgccaata ccagtagaaa cagacgaaga atccatgggt atggacagtt
    12061 ttccct
//
