LOCUS       sequence-383993-       11759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual gRNA vector targeting centromere-proximal locations on 
            Chromosome 7p and 7q in a third generation Cas9 backbone with GFP.
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: Chr7_Centromere-Targeting_gRNA
FEATURES             Location/Qualifiers
     source          1..11759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..340
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      360..369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             369..392
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             399..419
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             444..4544
                     /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             444..4544
                     /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             4545..4592
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4608..4664
                     /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(4665..4667,4668..4670,4671..5384)
                     /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(4665..4667,4668..4670,4671..5381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4971..4990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5318..5339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5400..5988
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5453..5473)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5871..5882)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6060..6293
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6371..6492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6408..6427)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6462..6481
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6532..6667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6687..6706)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6688..6706)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6716..6733)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6716..6732)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6725..6747)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6874..7329
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6961..6980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7171..7192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7355..7459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7460..7528,7529..8320)
                     /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(7678..7697)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8491..9079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8980..8999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9233..9250
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9367..9388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9403..9408,9409..9426,9427..9433)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9441..9457
                     /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     9446..9468
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9465..9481
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9465..9481
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9500..9520
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9502..9520
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9548..9774
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9775..9955
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10002..10127
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10620..10853
                     /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(11447..11695)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(11505..11524)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(11675..11695)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    join(11702..11759,1..60)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
       61 cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgagtg gctccggtgc
      121 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      181 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      241 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      301 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg
      361 ccaccatgga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg
      421 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca
      481 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
      541 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
      601 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
      661 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
      721 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
      781 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
      841 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
      901 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
      961 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     1021 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     1081 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     1141 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     1201 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     1261 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     1321 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     1381 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     1441 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     1501 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     1561 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     1621 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     1681 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     1741 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     1801 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     1861 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     1921 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     1981 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     2041 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     2101 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     2161 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     2221 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     2281 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     2341 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     2401 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     2461 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     2521 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     2581 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     2641 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     2701 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     2761 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     2821 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     2881 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     2941 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
     3001 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     3061 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
     3121 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     3181 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     3241 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
     3301 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     3361 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     3421 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
     3481 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
     3541 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
     3601 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
     3661 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
     3721 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
     3781 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
     3841 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
     3901 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
     3961 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
     4021 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
     4081 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
     4141 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
     4201 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
     4261 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
     4321 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
     4381 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
     4441 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
     4501 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaagcgt cctgctgcta
     4561 ctaagaaagc tggtcaagct aagaaaaaga aagctagcgg cagcggcgcc accaacttca
     4621 gcctgctgaa gcaggccggc gacgtggagg agaaccccgg ccccatggtg agcaagggcg
     4681 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4741 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4801 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4861 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     4921 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4981 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     5041 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     5101 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     5161 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     5221 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     5281 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     5341 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtagacgcgt taagtcgaca
     5401 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5461 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5521 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5581 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5641 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5701 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5761 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5821 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5881 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5941 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     6001 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     6061 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     6121 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6181 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6241 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta
     6301 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     6361 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6421 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6481 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     6541 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6601 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6661 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     6721 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     6781 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     6841 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     6901 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6961 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     7021 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7081 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7141 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7201 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7261 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     7321 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7381 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7441 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7501 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7561 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7621 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7681 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7741 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7801 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7861 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7921 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7981 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     8041 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     8101 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8161 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8221 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8281 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8341 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8401 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8461 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     8521 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8581 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     8641 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8701 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8761 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8821 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     8881 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8941 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     9001 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     9061 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     9121 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     9181 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     9241 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     9301 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     9361 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     9421 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     9481 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     9541 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     9601 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     9661 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     9721 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     9781 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     9841 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     9901 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     9961 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
    10021 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
    10081 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
    10141 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
    10201 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
    10261 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
    10321 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
    10381 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
    10441 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
    10501 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
    10561 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
    10621 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
    10681 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
    10741 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
    10801 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
    10861 tggggttgct ctggaaaact cgccatttgt ctcaagatct agaattcaaa aaaagcaccg
    10921 actcggtgcc actttgctgt ttccagcaaa gttgataacg gactagcctt atttaaactt
    10981 gctatgctgt ttccagcata gctcttaaac attggcttcc aattgaatca cgataaacat
    11041 ggcctctcaa gaactatata aagctaagaa tagaaaaaat tttacagtta tggtgactaa
    11101 actgtaaccc ttttttaagt ttcaaagtat ataataagtt atgacctttt gtaccacaca
    11161 cattatcatc atatagtttg attagggcta agtttaatag tctccataac cttattacat
    11221 catatatgca aatgaaggga accatgggaa attggcccta ggcccgcccg acctctgcgc
    11281 gtctgtctag tggactgcgc gggcagacaa gctcggtcac gcaaaaaaag caccgactcg
    11341 gtgccacttt gctgtttcca gcaaagttga taacggacta gccttatttc aacttgctat
    11401 gcactcttgt gcatagctct gaaaccaggt ctgttggagt ttactcggtg tttcgtcctt
    11461 tccacaagat atataaagcc aagaaatcga aatactttca agttacggta agcatatgat
    11521 agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac
    11581 gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcta attatctctc
    11641 taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcccaca
    11701 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacata
//
