LOCUS       sequence-384547-       11759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual gRNAs for deletion of OR6K6 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: OR6K6_Deletion_gRNA1
FEATURES             Location/Qualifiers
     source          1..11759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     404..421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    538..559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(574..579,580..597,598..604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..628
                     /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     617..639
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     636..652
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     636..652
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     671..691
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        673..691
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        719..945
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             946..1126
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1173..1298
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1791..2024
                     /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(2618..2866)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2676..2695)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2846..2866)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    2873..2990
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3059..3270
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3290..3299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3299..3322
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3329..3349
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3374..7474
                     /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             3374..7474
                     /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             7475..7522
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7538..7594
                     /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(7595..7597,7598..7600,7601..8314)
                     /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(7595..7597,7598..7600,7601..8311)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7901..7920)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8248..8269
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8330..8918
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8383..8403)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8801..8812)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8990..9223
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9301..9422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9338..9357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9392..9411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9462..9597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9617..9636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9618..9636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9646..9663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9646..9662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9655..9677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9804..10259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9891..9910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10101..10122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10285..10389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10390..10458,10459..11250)
                     /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(10608..10627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11421..11759,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      541 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      601 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      661 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
      721 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
      781 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
      841 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
      901 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
      961 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1021 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1081 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1141 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1201 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1261 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1321 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1381 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1441 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1501 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1561 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1621 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1681 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1741 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1801 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1861 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1921 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     1981 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2041 tctggaaaac tcgccatttg tctcaagatc tagaattcaa aaaaagcacc gactcggtgc
     2101 cactttgctg tttccagcaa agttgataac ggactagcct tatttaaact tgctatgctg
     2161 tttccagcat agctcttaaa cctgtggtac aatgtggggc acgataaaca tggcctctca
     2221 agaactatat aaagctaaga atagaaaaaa ttttacagtt atggtgacta aactgtaacc
     2281 cttttttaag tttcaaagta tataataagt tatgaccttt tgtaccacac acattatcat
     2341 catatagttt gattagggct aagtttaata gtctccataa ccttattaca tcatatatgc
     2401 aaatgaaggg aaccatggga aattggccct aggcccgccc gacctctgcg cgtctgtcta
     2461 gtggactgcg cgggcagaca agctcggtca cgcaaaaaaa gcaccgactc ggtgccactt
     2521 tgctgtttcc agcaaagttg ataacggact agccttattt caacttgcta tgcactcttg
     2581 tgcatagctc tgaaaccagt ggtctgctat acctaccggt gtttcgtcct ttccacaaga
     2641 tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt
     2701 taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat
     2761 tttgtactaa tatctttgtg tttacagtca aattaattct aattatctct ctaacagcct
     2821 tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcccac aattttaaaa
     2881 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2941 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     3001 acagggacag cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg
     3061 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     3121 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     3181 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     3241 tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga
     3301 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
     3361 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     3421 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     3481 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     3541 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     3601 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     3661 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     3721 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     3781 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     3841 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     3901 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     3961 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     4021 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     4081 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     4141 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     4201 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     4261 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     4321 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     4381 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     4441 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     4501 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     4561 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     4621 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     4681 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     4741 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     4801 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     4861 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     4921 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     4981 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     5041 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     5101 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     5161 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     5221 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     5281 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     5341 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     5401 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     5461 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     5521 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     5581 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     5641 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     5701 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     5761 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     5821 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     5881 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     5941 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     6001 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     6061 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     6121 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     6181 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     6241 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     6301 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     6361 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     6421 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     6481 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     6541 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     6601 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     6661 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     6721 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     6781 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     6841 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     6901 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     6961 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     7021 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     7081 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     7141 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     7201 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     7261 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     7321 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     7381 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     7441 gacacggatc gacctgtctc agctgggagg cgacaagcgt cctgctgcta ctaagaaagc
     7501 tggtcaagct aagaaaaaga aagctagcgg cagcggcgcc accaacttca gcctgctgaa
     7561 gcaggccggc gacgtggagg agaaccccgg ccccatggtg agcaagggcg aggagctgtt
     7621 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     7681 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     7741 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     7801 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     7861 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     7921 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     7981 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     8041 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     8101 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     8161 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     8221 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     8281 gatcactctc ggcatggacg agctgtacaa gtagacgcgt taagtcgaca atcaacctct
     8341 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     8401 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     8461 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     8521 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     8581 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     8641 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     8701 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     8761 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     8821 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     8881 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
     8941 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     9001 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     9061 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     9121 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     9181 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
     9241 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     9301 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     9361 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     9421 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     9481 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     9541 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     9601 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     9661 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     9721 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     9781 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     9901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     9961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    10021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    10081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    10141 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    10201 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
    10261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
    10321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
    10381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
    10441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
    10501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
    10561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
    10621 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
    10681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
    10741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
    10801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
    10861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
    10921 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
    10981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
    11041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
    11101 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
    11161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
    11221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
    11281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
    11341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
    11401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
    11461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
    11521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
    11581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
    11641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
    11701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacaca
//
