LOCUS       sequence-404975-       11759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual gRNA plasmid for UCK2 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11759)
  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 11759)
  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. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: UCK2_Deletion_gRNA_2
FEATURES             Location/Qualifiers
     source          1..11759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      928..1516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1417..1436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1670..1687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1840..1845,1846..1863,1864..1870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1878..1894
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1883..1905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1902..1918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1937..1957
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1939..1957
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1985..2211
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2212..2392
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2439..2564
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3057..3290
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        complement(3884..4132)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(3942..3961)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4112..4132)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    4139..4256
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4325..4536
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4556..4565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4565..4588
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4595..4615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4640..8740
                     /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             4640..8740
                     /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             8741..8788
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8804..8860
                     /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(8861..8863,8864..8866,8867..9580)
                     /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(8861..8863,8864..8866,8867..9577)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9167..9186)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9514..9535
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    9596..10184
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9649..9669)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10067..10078)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10256..10489
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10567..10688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10604..10623)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10658..10677
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10728..10863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10883..10902)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10884..10902)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10912..10929)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10912..10928)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10921..10943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11070..11525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11157..11176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11367..11388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11551..11655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11656..11724,11725..11759,1..757)
                     /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"
ORIGIN
        1 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     1981 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
     2041 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
     2101 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
     2161 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
     2221 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     2281 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     2341 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     2401 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     2461 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     2521 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     2581 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     2641 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     2701 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     2761 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     2821 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     2881 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     2941 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     3001 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     3061 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     3121 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     3181 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     3241 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     3301 ggttgctctg gaaaactcgc catttgtctc aagatctaga attcaaaaaa agcaccgact
     3361 cggtgccact ttgctgtttc cagcaaagtt gataacggac tagccttatt taaacttgct
     3421 atgctgtttc cagcatagct cttaaactgg tctagtggat gttctaccga taaacatggc
     3481 ctctcaagaa ctatataaag ctaagaatag aaaaaatttt acagttatgg tgactaaact
     3541 gtaacccttt tttaagtttc aaagtatata ataagttatg accttttgta ccacacacat
     3601 tatcatcata tagtttgatt agggctaagt ttaatagtct ccataacctt attacatcat
     3661 atatgcaaat gaagggaacc atgggaaatt ggccctaggc ccgcccgacc tctgcgcgtc
     3721 tgtctagtgg actgcgcggg cagacaagct cggtcacgca aaaaaagcac cgactcggtg
     3781 ccactttgct gtttccagca aagttgataa cggactagcc ttatttcaac ttgctatgca
     3841 ctcttgtgca tagctctgaa accgtgcgac tctctagtga agcggtgttt cgtcctttcc
     3901 acaagatata taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt
     3961 ccattttaaa acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc
     4021 acgtattttg tactaatatc tttgtgttta cagtcaaatt aattctaatt atctctctaa
     4081 cagccttgta tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcccacaatt
     4141 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     4201 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     4261 tttattacag ggacagcaga gatccacttt ggcgccggct cgagtggctc cggtgcccgt
     4321 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     4381 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     4441 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     4501 gttctttttc gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg gatccgccac
     4561 catggattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
     4621 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     4681 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     4741 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     4801 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     4861 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     4921 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     4981 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     5041 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     5101 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     5161 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     5221 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     5281 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     5341 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     5401 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     5461 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     5521 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     5581 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     5641 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     5701 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     5761 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     5821 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     5881 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     5941 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     6001 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     6061 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     6121 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     6181 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     6241 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     6301 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     6361 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     6421 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     6481 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     6541 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     6601 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     6661 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     6721 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     6781 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     6841 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     6901 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     6961 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     7021 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     7081 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     7141 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     7201 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     7261 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     7321 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     7381 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     7441 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     7501 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     7561 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     7621 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     7681 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     7741 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     7801 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     7861 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     7921 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     7981 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     8041 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     8101 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     8161 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     8221 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     8281 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     8341 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     8401 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     8461 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     8521 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     8581 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     8641 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     8701 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aagcgtcctg ctgctactaa
     8761 gaaagctggt caagctaaga aaaagaaagc tagcggcagc ggcgccacca acttcagcct
     8821 gctgaagcag gccggcgacg tggaggagaa ccccggcccc atggtgagca agggcgagga
     8881 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     8941 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     9001 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     9061 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     9121 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     9181 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     9241 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     9301 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     9361 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     9421 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     9481 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     9541 cgccgggatc actctcggca tggacgagct gtacaagtag acgcgttaag tcgacaatca
     9601 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     9661 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     9721 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     9781 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     9841 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     9901 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     9961 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
    10021 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
    10081 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    10141 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
    10201 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
    10261 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
    10321 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
    10381 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
    10441 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tacgtatagt
    10501 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
    10561 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    10621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    10681 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
    10741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
    10801 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    10861 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
    10921 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
    10981 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
    11041 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
    11101 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
    11161 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
    11221 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
    11281 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
    11341 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
    11401 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
    11461 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
    11521 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
    11581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
    11641 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
    11701 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctga
//
