LOCUS       sequence-273968-        9251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA to delete a nucleation site near Lmx1b. Use with 
            LMX1B-gRNA-C-del.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9251)
  AUTHORS   Oksuz O, Narendra V, Lee CH, Descostes N, LeRoy G, Raviram R, 
            Blumenberg L, Karch K, Rocha PP, Garcia BA, Skok JA, Reinberg D
  TITLE     Capturing the Onset of PRC2-Mediated Repressive Domain Formation.
  JOURNAL   Mol Cell. 2018 Jun 21;70(6):1149-1162.e5. doi: 
            10.1016/j.molcel.2018.05.023.
  PUBMED    29932905
REFERENCE   2  (bases 1 to 9251)
  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: 
            "10.1016/j.molcel.2018.05"; journalName: "Mol Cell"; date: 
            "2018-06-21- 21"; volume: "70"; issue: "6"; pages: "1149-1162.e5"
COMMENT     Sequence Label: LMX1B-gRNA-N-del
FEATURES             Location/Qualifiers
     source          1..9251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          127..354
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      366..375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             375..440
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             447..467
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             492..4592
                     /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             492..4592
                     /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             4593..4640
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4656..4709
                     /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             join(4710..4712,4713..5423)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4710..4712,4713..5423)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5013..5032)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5360..5381
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5451..5468)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5457..5664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5673..5813
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5673..5802
                     /label=AAV2 ITR
     rep_origin      5888..6343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5975..5994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6185..6206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6360..6379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6479..6501
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6539..6557)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6625..6729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6730..6798,6799..7590)
                     /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(6948..6967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7761..8349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8250..8269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8411..8659
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8411..8431
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8582..8601
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8681..8756
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8853..9138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      181 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      241 tctcctccgg gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt
      301 ggggtattaa tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga
      361 ccggtgccac catggactat aaggaccacg acggagacta caaggatcat gatattgatt
      421 acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag
      481 tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
      541 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
      601 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
      661 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
      721 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
      781 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
      841 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
      901 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
      961 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     1021 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     1081 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     1141 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     1201 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     1261 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     1321 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     1381 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     1441 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     1501 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     1561 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     1621 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     1681 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     1741 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     1801 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     1861 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     1921 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     1981 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     2041 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     2101 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     2161 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     2221 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     2281 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     2341 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     2401 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     2461 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     2521 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     2581 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     2641 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     2701 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     2761 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     2821 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     2881 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     2941 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     3001 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     3061 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     3121 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     3181 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     3241 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     3301 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     3361 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     3421 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     3481 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     3541 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     3601 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     3661 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     3721 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     3781 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     3841 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     3901 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     3961 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     4021 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     4081 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     4141 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     4201 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     4261 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     4321 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     4381 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     4441 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     4501 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     4561 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
     4621 gccaggcaaa aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat
     4681 gcggtgacgt cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg
     4741 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     4801 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4861 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     4921 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4981 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     5041 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     5101 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     5161 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     5221 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     5281 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     5341 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     5401 tcggcatgga cgagctgtac aaggaattct aactagagct cgctgatcag cctcgactgt
     5461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5641 agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac
     5701 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     5761 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     5821 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     5881 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5941 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     6001 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6061 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     6121 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6181 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     6241 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     6301 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     6361 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6421 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6481 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6541 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6601 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6661 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6721 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6781 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6841 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6901 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6961 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7021 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7081 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7141 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7201 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7261 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7321 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7381 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7441 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7501 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7561 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7621 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7681 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7741 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7801 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7861 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7921 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7981 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8041 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8101 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8161 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8221 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8281 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8341 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8401 gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     8461 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     8521 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     8581 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     8641 gtggaaagga cgaaacaccg cctccaggtt tggttccagc gttttagagc tagaaatagc
     8701 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     8761 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     8821 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
     8881 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     8941 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     9001 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     9061 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     9121 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     9181 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     9241 ggggcggggg g
//
