LOCUS       sequence-383924-        9682 bp    DNA     circular SYN 15-APR-2025
DEFINITION  gRNA in a third generation Cas9 vector with GFP, targeting region 
            immediately upstream of MDM4, to be used with 
            MDM4_Deletion_Downstream_gRNA_1/2 for MDM4 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9682)
  AUTHORS   Girish V, Lakhani AA, Thompson SL, Scaduto CM, Brown LM, Hagenson 
            RA, Sausville EL, Mendelson BE, Kandikuppa PK, Lukow DA, Yuan ML, 
            Stevens EC, Lee SN, Schukken KM, Akalu SM, Vasudevan A, Zou C, 
            Salovska B, Li W, Smith JC, Taylor AM, Martienssen RA, Liu Y, Sun R,
            Sheltzer JM
  TITLE     Oncogene-like addiction to aneuploidy in human cancers.
  JOURNAL   Science. 2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521.
  PUBMED    37410869
REFERENCE   2  (bases 1 to 9682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: MDM4_Deletion_Upstream_gRNA_1
FEATURES             Location/Qualifiers
     source          1..9682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(3..250)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(60..79)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(230..250)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    257..374
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        443..654
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      674..683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             683..706
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             713..733
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             758..4858
                     /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             758..4858
                     /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             4859..4906
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4922..4978
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(4979..4981,4982..4984,4985..5698)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4979..4981,4982..4984,4985..5695)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5285..5304)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5632..5653
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5714..6302
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5767..5787)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6185..6196)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6374..6607
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6685..6806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6722..6741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6776..6795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6846..6981
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7001..7020)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7002..7020)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7030..7047)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7030..7046)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7039..7061)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7188..7643
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7275..7294)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7485..7506
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7669..7773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7774..7842,7843..8634)
                     /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(7992..8011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8805..9393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9294..9313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9547..9564
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgtgtttcg tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta
       61 cggtaagcat atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa
      121 attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa
      181 ttctaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa
      241 ataggccctc ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
      301 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
      361 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg cgccggctcg
      421 agtggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
      481 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
      541 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
      601 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtgtcg
      661 tgacgcggga tccgccacca tggattacaa agacgatgac gataagatgg ccccaaagaa
      721 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
      781 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
      841 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
      901 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
      961 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1021 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1081 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1141 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     1201 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     1261 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1321 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     1381 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     1441 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     1501 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     1561 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     1621 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     1681 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     1741 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     1801 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     1861 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     1921 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     1981 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2041 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2101 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     2161 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     2221 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     2281 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     2341 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     2401 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     2461 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     2521 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     2581 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     2641 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     2701 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     2761 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     2821 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     2881 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     2941 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3001 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     3061 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     3121 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     3181 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     3241 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     3301 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     3361 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     3421 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     3481 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     3541 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     3601 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     3661 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     3721 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     3781 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     3841 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     3901 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     3961 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4021 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     4081 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     4141 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     4201 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     4261 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4321 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     4381 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     4441 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     4501 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     4561 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     4621 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     4681 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     4741 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     4801 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     4861 gcgtcctgct gctactaaga aagctggtca agctaagaaa aagaaagcta gcggcagcgg
     4921 cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc ccggccccat
     4981 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     5041 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     5101 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     5161 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     5221 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     5281 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     5341 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     5401 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     5461 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     5521 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5581 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5641 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagac
     5701 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5761 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5821 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5881 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5941 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6001 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6061 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     6121 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6181 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6241 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     6301 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6361 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     6421 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6481 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6541 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6601 tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6661 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6781 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6901 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6961 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     7021 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     7081 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     7141 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     7201 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7261 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7321 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7381 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7441 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7501 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     7561 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7621 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     7681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7861 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7921 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7981 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     8041 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     8101 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     8161 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8221 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8281 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     8341 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8401 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8461 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8521 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8581 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8641 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8701 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8761 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8821 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8881 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8941 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     9001 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     9061 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     9121 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9181 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9241 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9301 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9361 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9421 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9481 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9541 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9601 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     9661 ggcagtgagc gcaacgcaat ta
//
