LOCUS       sequence-384546-       11759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual gRNAs for deletion of OR2M4 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: OR2M4_Deletion_gRNA
FEATURES             Location/Qualifiers
     source          1..11759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(535..540,541..558,559..565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    573..589
                     /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     578..600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     597..613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     597..613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     632..652
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        634..652
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        680..906
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             907..1087
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1134..1259
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1752..1985
                     /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(2579..2827)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2637..2656)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2807..2827)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    2834..2951
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3020..3231
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3251..3260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3260..3283
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3290..3310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3335..7435
                     /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             3335..7435
                     /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             7436..7483
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7499..7555
                     /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(7556..7558,7559..7561,7562..8275)
                     /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(7556..7558,7559..7561,7562..8272)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7862..7881)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8209..8230
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8291..8879
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8344..8364)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8762..8773)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8951..9184
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9262..9383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9299..9318)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9353..9372
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9423..9558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9578..9597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9579..9597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9607..9624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9607..9623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9616..9638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9765..10220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9852..9871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10062..10083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10246..10350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10351..10419,10420..11211)
                     /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(10569..10588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11382..11759,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      541 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      601 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
      661 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
      721 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
      781 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
      841 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
      901 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      961 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1021 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1081 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     1141 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1201 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1261 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1321 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1381 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1441 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1501 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1561 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     1621 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1681 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1741 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1801 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1861 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1921 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1981 ctcctgggga tttggggttg ctctggaaaa ctcgccattt gtctcaagat ctagaattca
     2041 aaaaaagcac cgactcggtg ccactttgct gtttccagca aagttgataa cggactagcc
     2101 ttatttaaac ttgctatgct gtttccagca tagctcttaa accgggacat ctccattgaa
     2161 cacgataaac atggcctctc aagaactata taaagctaag aatagaaaaa attttacagt
     2221 tatggtgact aaactgtaac ccttttttaa gtttcaaagt atataataag ttatgacctt
     2281 ttgtaccaca cacattatca tcatatagtt tgattagggc taagtttaat agtctccata
     2341 accttattac atcatatatg caaatgaagg gaaccatggg aaattggccc taggcccgcc
     2401 cgacctctgc gcgtctgtct agtggactgc gcgggcagac aagctcggtc acgcaaaaaa
     2461 agcaccgact cggtgccact ttgctgtttc cagcaaagtt gataacggac tagccttatt
     2521 tcaacttgct atgcactctt gtgcatagct ctgaaactcg gacaagtggg tcatttgcgg
     2581 tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg
     2641 taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt
     2701 attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc
     2761 taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata
     2821 ggccctccca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     2881 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     2941 ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc cggctcgagt
     3001 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3061 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga
     3241 cgcgggatcc gccaccatgg attacaaaga cgatgacgat aagatggccc caaagaagaa
     3301 gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga
     3361 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     3421 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     3481 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     3541 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     3601 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     3661 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     3721 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     3781 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     3841 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     3901 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     3961 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     4021 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     4081 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     4141 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     4201 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     4261 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     4321 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     4381 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     4441 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     4501 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     4561 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     4621 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     4681 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     4741 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     4801 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     4861 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     4921 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     4981 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     5041 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     5101 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     5161 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     5221 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     5281 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     5341 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     5401 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     5461 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     5521 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     5581 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     5641 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     5701 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     5761 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     5821 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     5881 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     5941 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     6001 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     6061 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     6121 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     6181 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     6241 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     6301 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     6361 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     6421 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     6481 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     6541 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     6601 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     6661 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     6721 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     6781 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     6841 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     6901 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     6961 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     7021 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     7081 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     7141 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     7201 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     7261 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     7321 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     7381 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg
     7441 tcctgctgct actaagaaag ctggtcaagc taagaaaaag aaagctagcg gcagcggcgc
     7501 caccaacttc agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccatggt
     7561 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     7621 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     7681 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     7741 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     7801 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     7861 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     7921 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     7981 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     8041 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     8101 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     8161 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     8221 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtagacgcg
     8281 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     8341 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     8401 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     8461 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     8521 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     8581 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     8641 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     8701 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     8761 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     8821 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     8881 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     8941 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     9001 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     9061 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     9121 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     9181 agcagtacgt atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     9241 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     9301 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     9361 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     9421 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     9481 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     9541 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     9601 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     9661 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     9721 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     9781 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     9841 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     9901 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     9961 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
    10021 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
    10081 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
    10141 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
    10201 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
    10261 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
    10321 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
    10381 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
    10441 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
    10501 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
    10561 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
    10621 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
    10681 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
    10741 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
    10801 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
    10861 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
    10921 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
    10981 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
    11041 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
    11101 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
    11161 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
    11221 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
    11281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
    11341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
    11401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
    11461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
    11521 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
    11581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
    11641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
    11701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagata
//
