LOCUS       sequence-429354-       14197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An all-in-one system for on-target efficiency analysis of CRISPR 
            guide RNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14197)
  AUTHORS   Lin D, Najam SS, Liu Y, Murgia N, Vinnikov IA
  TITLE     Noodles, the all-in-one system for on-target efficiency analysis of 
            CRISPR guide RNAs.
  JOURNAL   MethodsX. 2023 Nov 15;12:102481. doi: 10.1016/j.mex.2023.102481. 
            eCollection 2024 Jun.
  PUBMED    38162150
REFERENCE   2  (bases 1 to 14197)
  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: "MethodsX.";
            date: "2023-11-15"; volume: "12:102481. doi"; pages: " 
            10.1016/j.mex.2023.102481. eCollection 2024 Jun"
COMMENT     Sequence Label: Noodles Plasmid
FEATURES             Location/Qualifiers
     source          1..14197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4..195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(27..47)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      212..347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      406..415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             412..1344
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1396..2187
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1447..1466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2057..2076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2251..2299
                     /note="synthetic polyadenylation signal"
     promoter        2455..2559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2560..2628,2629..3420)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2778..2797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3578..4166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3658..3677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4282..4565
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        4679..4982
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4983..5186
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5137..5157
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5183..5207
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5305..5314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    7376..7497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7387..7406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7441..7460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        7513..7761
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7513..7533
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7684..7703
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9643..9718
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        9780..9991
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      10010..10019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10016..14119
                     /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             10016..14119
                     /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             14120..14167
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             14168..14191
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
       61 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
      121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      181 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
      241 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
      301 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
      361 aaagcttgat tcttctgaca caacagtctc gaaccaaagg ctggagccac catggcttcc
      421 aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc
      481 tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc
      541 gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc
      601 gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag
      661 tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct
      721 tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct
      781 tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct
      841 gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc
      901 gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag
      961 accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg
     1021 gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc
     1081 cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac
     1141 cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc
     1201 aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc
     1261 ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg
     1321 gagcgcgtgc tgaagaacga gcagaccggt ggtgggagcg gaggtggcgg atcaggtggc
     1381 ggaggctccg gagggattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     1441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     1501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     1561 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     1621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     1681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     1741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     1801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     1861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     1921 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     1981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     2041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     2101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     2161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     2221 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     2281 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc
     2341 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     2401 gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2461 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2521 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2581 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2641 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2701 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2761 agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2821 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2881 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2941 agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc
     3001 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3061 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3121 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3181 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3241 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3301 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3361 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3421 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     3481 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3541 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3601 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3661 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3721 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3781 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3841 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3901 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3961 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4021 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4081 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4141 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     4201 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4261 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     4321 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     4381 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     4441 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     4501 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     4561 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatccta
     4621 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4681 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4741 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4801 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4861 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4921 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4981 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
     5041 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5101 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
     5161 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta
     5221 ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgggcctcgg cggccaagct
     5281 tggcaatccg gtactgttgg taaagccacc atggaagatg ccaaaaacat taagaagggc
     5341 ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg
     5401 aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat cgaggtggac
     5461 attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat
     5521 gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg
     5581 cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac
     5641 gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag
     5701 aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc
     5761 atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc
     5821 catttgccac ccggcttcaa cgagtacgac ttcgtgcccg aataatgagg cgcgaccatc
     5881 ttcttcaagg acgacggcgc gcctgggatc ctgcaggtgt gcagcgagaa tagcttgcag
     5941 ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg ctgtggcccc agctaacgac
     6001 atctacaacg agcgcgagct gctgaacagc atgggcatca gccagcccac cgtcgtattc
     6061 gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa agaagctacc gatcatacaa
     6121 aagatcatca tcatggatag caagaccgac taccagggct tccaaagcat gtacaccttc
     6181 gtgacttccc atttgccacc cggcttcaac gagtacgact tcgtgcccga gagcttcgac
     6241 cgggacaaaa ccatcgccct gatcatgaac agtagtggca gtaccggatt gcccaagggc
     6301 gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc
     6361 ggcaaccaga tcatccccga caccgctatc ctcagcgtgg tgccatttca ccacggcttc
     6421 ggcatgttca ccacgctggg ctacttgatc tgcggctttc gggtcgtgct catgtaccgc
     6481 ttcgaggagg agctattctt gcgcagcttg caagactata agattcaatc tgccctgctg
     6541 gtgcccacac tatttagctt cttcgctaag agcactctca tcgacaagta cgacctaagc
     6601 aacttgcacg agatcgccag cggcggggcg ccgctcagca aggaggtagg tgaggccgtg
     6661 gccaaacgct tccacctacc aggcatccgc cagggctacg gcctgacaga aacaaccagc
     6721 gccattctga tcacccccga aggggacgac aagcctggcg cagtaggcaa ggtggtgccc
     6781 ttcttcgagg ctaaggtggt ggacttggac accggtaaga cactgggtgt gaaccagcgc
     6841 ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct acgttaacaa ccccgaggct
     6901 acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg gcgacatcgc ctactgggac
     6961 gaggacgagc acttcttcat cgtggaccgg ctgaagagcc tgatcaaata caagggctac
     7021 caggtagccc cagccgaact ggagagcatc ctgctgcaac accccaacat cttcgacgcc
     7081 ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg
     7141 gaacacggta aaaccatgac cgagaaggag atcgtggact atgtggccag ccaggttaca
     7201 accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc
     7261 ggcaagttgg acgcccgcaa gatccgcgag attctcatta aggccaagaa gggcggcaag
     7321 atcgccgtgt aataattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
     7381 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     7441 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     7501 aacaacaatt gcgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     7561 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     7621 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     7681 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     7741 ttgtggaaag gacgaaacac cggagacggt tgtaaatgag cacacaaaat acacatgcta
     7801 aaatattata ttctatgacc tttataaaat caaccaaaat cttcttttta ataactttag
     7861 tatcaataat tagaattttt atgttccttt ttgcaaactt ttaataaaaa tgagcaaaat
     7921 aaaaaaacgc tagttttagt aactcgcgtt gttttcttca cctttaataa tagctactcc
     7981 accacttgtt cctaagcggt cagctcctgc ttcaatcatt ttttgagcat cttcaaatgt
     8041 tctaactcca ccagctgctt taactaaagc attgtcttta acaactgact tcattagttt
     8101 aacatcttca aatgttgcac ctgattttga aaatcctgtt gatgttttaa caaattctaa
     8161 tccagcttca acagctattt cacaagcttt catgatttct tcttttgtta ataaacaatt
     8221 ttccataata catttaacaa catgtgatcc agctgctttt tttacagctt tcatgtcttc
     8281 taaaactaat tcataatttt tgtcttttaa tgcaccaata tttaatacca tatcaatttc
     8341 tgttgcacca tctttaattg cttcagaaac ttcgaatgct tttgtagctg ttgtgcatgc
     8401 acctagagga aaacctacaa catttgttat tcctacattt gtgcctttta ataattcttt
     8461 acaatagctt gttcaatatg aattaacaca aactgttgca aaatcaaatt caattgcttc
     8521 atcacataat tgtttaattt cagctttcgt agcatcttgt tttaataatg tgtgatctat
     8581 atatttgttt agtttcattt tttctcctat atattcattt ttaattttaa ttctttaata
     8641 atttcgtcta ctttaacttt agcgttttga acagattcac caacacctat aaaataaatt
     8701 tttagtttag gttcagttcc acttgggcga acagcaaatc atgacttatc ttctaaataa
     8761 aattttagta agtcttgtcc tggcatatta tacattccat cgatgtagtc ttcaacatta
     8821 acaactttaa gtccagcaat ttgagttaag ggtgttgctc tcaatgattt cattaatggt
     8881 tcaattttta atttcttttc ttctggttta aaattcaagt ttaaagtgaa agtgtaatat
     8941 gcacccattt ctttaaataa atcttctaaa tagtctacta atgttttatt ttgtttttta
     9001 taaaatcaag cagcctctgc tattaatata gaagcttgta ttccatcttt atctctagct
     9061 gagtcatcaa ttacatatcc ataactttct tcataagcaa aaacaaaatt taatccgtta
     9121 tcttcttctt tagcaatttc tctacccatt catttaaatc cagttaaagt ttttacaata
     9181 ttaactccat atttttcatg agcgattcta tcacccaaat cacttgttac aaaacttgaa
     9241 tatagagccg gattttttgg aatgctattt aagcgtttta gatttgataa ttttcaatca
     9301 attaaaattg gtcctgtttg atttccatct aatcttacaa aatgaccatc atgttttatt
     9361 gccattccaa atctgtcagc atctgggtca ttcataataa taatatctgc atcatgttta
     9421 ataccatatt caagcggtat ttttcatgca ggatcaaatt ctggatttgg atttacaaca
     9481 tttttaaatg tttcatcttc aaatgcatgc tcttcaacct caataacgtt atatcctgat
     9541 tcacgtaata tttttggggt aaatttagtt cctgttccat taactgcgct aaaaataatt
     9601 tttaaatctt ttttagcttc ttgctctttt ttgtacgtct ctgttttaga gctagaaata
     9661 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     9721 ttttgaattc gctagctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
     9781 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgatc
     9841 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     9901 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     9961 ttttcgcaac gggtttgccg ccagaacaca ggaccggttc tagagcgctg ccaccatgga
    10021 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
    10081 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
    10141 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
    10201 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
    10261 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
    10321 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
    10381 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
    10441 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
    10501 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
    10561 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
    10621 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
    10681 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
    10741 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
    10801 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
    10861 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
    10921 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
    10981 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
    11041 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
    11101 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
    11161 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
    11221 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
    11281 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
    11341 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
    11401 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
    11461 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
    11521 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
    11581 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
    11641 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
    11701 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
    11761 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
    11821 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
    11881 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
    11941 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
    12001 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
    12061 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
    12121 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
    12181 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
    12241 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
    12301 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
    12361 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
    12421 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
    12481 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
    12541 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
    12601 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
    12661 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
    12721 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
    12781 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
    12841 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
    12901 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
    12961 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
    13021 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
    13081 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
    13141 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
    13201 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
    13261 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
    13321 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
    13381 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
    13441 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
    13501 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
    13561 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
    13621 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
    13681 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
    13741 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
    13801 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
    13861 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
    13921 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
    13981 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
    14041 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
    14101 gtctcagctg ggaggcgaca agcgacctgc cgccacaaag aaggctggac aggctaagaa
    14161 gaagaaagat tacaaagacg atgacgataa ggggtac
//
