LOCUS       sequence-383939-       11344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA targeting centromere-proximal location on Chromosome 8q in a 
            third generation Cas9 backbone with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11344)
  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 11344)
  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: Chr8q_Centromere-Targeting_gRNA
FEATURES             Location/Qualifiers
     source          1..11344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /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(1490..1738)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1548..1567)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1718..1738)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    1745..1862
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1931..2142
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2162..2171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2171..2194
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2201..2221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2246..6346
                     /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             2246..6346
                     /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             6347..6394
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6410..6466
                     /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(6467..6469,6470..6472,6473..7186)
                     /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(6467..6469,6470..6472,6473..7183)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6773..6792)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7120..7141
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7202..7790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7255..7275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7673..7684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7862..8095
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8173..8294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8210..8229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8264..8283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8334..8469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8489..8508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8490..8508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8518..8535)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8518..8534)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8527..8549)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8676..9131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8763..8782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8973..8994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9157..9261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9262..9330,9331..10122)
                     /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(9480..9499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10293..10881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10782..10801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11035..11052
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11169..11190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11205..11210,11211..11228,11229..11235)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11243..11259
                     /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     11248..11270
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11267..11283
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11267..11283
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11302..11322
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11304..11322
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactcg ccatttgtct caagatctag aattcaaaaa aagcaccgac
     1381 tcggtgccac tttgctgttt ccagcaaagt tgataacgga ctagccttat ttaaacttgc
     1441 tatgctgttt ccagcatagc tcttaaaccg tcttttgtaa cccacctgcg gtgtttcgtc
     1501 ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat
     1561 gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc
     1621 tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt ctaattatct
     1681 ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctccc
     1741 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     1801 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     1861 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag tggctccggt
     1921 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     1981 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2041 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2101 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc
     2161 cgccaccatg gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
     2221 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
     2281 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     2341 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     2401 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     2461 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     2521 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     2581 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     2641 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     2701 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     2761 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     2821 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     2881 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     2941 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     3001 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     3061 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     3121 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     3181 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     3241 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     3301 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     3361 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     3421 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     3481 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     3541 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     3601 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     3661 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     3721 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     3781 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     3841 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     3901 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     3961 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     4021 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     4081 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     4141 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     4201 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     4261 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     4321 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     4381 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     4441 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     4501 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     4561 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     4621 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     4681 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     4741 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     4801 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     4861 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     4921 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     4981 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     5041 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     5101 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     5161 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     5221 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     5281 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     5341 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     5401 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     5461 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     5521 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     5581 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     5641 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     5701 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     5761 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     5821 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     5881 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     5941 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     6001 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     6061 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     6121 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     6181 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     6241 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     6301 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaagc gtcctgctgc
     6361 tactaagaaa gctggtcaag ctaagaaaaa gaaagctagc ggcagcggcg ccaccaactt
     6421 cagcctgctg aagcaggccg gcgacgtgga ggagaacccc ggccccatgg tgagcaaggg
     6481 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     6541 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     6601 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     6661 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     6721 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     6781 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     6841 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     6901 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     6961 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     7021 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     7081 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     7141 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtagacgc gttaagtcga
     7201 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     7261 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     7321 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     7381 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     7441 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     7501 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     7561 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     7621 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     7681 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     7741 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     7801 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     7861 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     7921 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7981 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     8041 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     8101 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     8161 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     8221 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8281 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     8341 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     8401 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     8461 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     8521 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     8581 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     8641 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     8701 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     8761 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     8821 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     8881 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     8941 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     9001 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     9061 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     9121 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     9181 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     9241 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     9301 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     9361 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     9421 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     9481 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     9541 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     9601 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     9661 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     9721 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     9781 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     9841 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     9901 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9961 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
    10021 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
    10081 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
    10141 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
    10201 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
    10261 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
    10321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    10381 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    10441 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    10501 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    10561 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    10621 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    10681 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    10741 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    10801 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    10861 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    10921 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    10981 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    11041 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
    11101 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
    11161 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
    11221 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
    11281 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
    11341 caag
//
