LOCUS       sequence-467526-        9177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes CRISPR gRNA for cleaving upstream of insertion site at C 
            terminus of mouse Rpl31.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9177)
  AUTHORS   Zhang Z, Xu A, Bai Y, Chen Y, Cates K, Kerr C, Bermudez A, Susanto 
            TT, Wysong K, Garcia Marques FJ, Nolan GP, Pitteri S, Barna M
  TITLE     A subcellular map of translational machinery composition and 
            regulation at the single-molecule level.
  JOURNAL   Science. 2025 Mar 7;387(6738):eadn2623. doi: 
            10.1126/science.adn2623. Epub 2025 Mar 7.
  PUBMED    40048539
REFERENCE   2  (bases 1 to 9177)
  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: "Science."; 
            date: "2025-03-7"; volume: "387(6738):eadn2623. doi"; pages: " 
            10.1126/science.adn2623. Epub 2025 Mar 7"
COMMENT     Sequence Label: PX459_L31_CRISPR_upstream_sgRNA
FEATURES             Location/Qualifiers
     source          1..9177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             573..620
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             636..689
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             690..1289
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(690..709)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1186..1206
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1317..1334)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1323..1530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1539..1679
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1539..1668
                     /label=AAV2 ITR
     rep_origin      1754..2209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1841..1860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2051..2072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2226..2245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2345..2367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2405..2423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2491..2595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2596..2664,2665..3456)
                     /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(2814..2833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3627..4215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4116..4135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4277..4525
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4277..4297
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4448..4467
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4547..4622
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        4719..5004
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        5006..5283
                     /label=chicken beta-actin promoter
     intron          5284..5511
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5523..5532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5532..5597
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5604..5624
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5649..9177,1..572)
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(5649..9177,1..572)
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
       61 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
      121 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
      181 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
      241 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
      301 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
      361 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
      421 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
      481 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
      541 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
      601 gccaggcaaa aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat
      661 gcggtgacgt cgaggagaat cctggcccaa tgaccgagta caagcccacg gtgcgcctcg
      721 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
      781 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
      841 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
      901 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
      961 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     1021 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     1081 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     1141 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     1201 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     1261 ggtgcatgac ccgcaagccc ggtgcctgag aattctaact agagctcgct gatcagcctc
     1321 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1381 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1441 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1501 ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt
     1561 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     1621 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     1681 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     1741 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1801 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     1861 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1921 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     1981 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2041 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     2101 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     2161 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     2221 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2281 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2341 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2401 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2461 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2521 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2581 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2641 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2701 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2761 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2821 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2881 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2941 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3001 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3061 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3121 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3181 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3241 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3301 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3361 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3421 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3481 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3541 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3601 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3661 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3721 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3781 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3841 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3901 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3961 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4021 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4081 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4141 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4201 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4261 ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     4321 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     4381 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     4441 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     4501 tatcttgtgg aaaggacgaa acaccgacgt tgaccgtctg tagatcgttt tagagctaga
     4561 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     4621 gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc
     4681 gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat
     4741 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata
     4801 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4861 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     4921 gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4981 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     5041 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     5101 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     5161 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     5221 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     5281 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     5341 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     5401 cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg
     5461 gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg
     5521 gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca
     5581 aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc
     5641 cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg
     5701 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     5761 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     5821 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     5881 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     5941 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     6001 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     6061 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     6121 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     6181 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     6241 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     6301 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     6361 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     6421 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     6481 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     6541 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     6601 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     6661 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     6721 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     6781 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     6841 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     6901 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     6961 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     7021 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     7081 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     7141 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     7201 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     7261 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     7321 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     7381 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     7441 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     7501 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     7561 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     7621 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     7681 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     7741 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     7801 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     7861 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     7921 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     7981 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     8041 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     8101 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     8161 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     8221 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     8281 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     8341 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     8401 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     8461 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     8521 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     8581 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     8641 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     8701 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     8761 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     8821 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     8881 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     8941 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     9001 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     9061 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     9121 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatc
//
