LOCUS       sequence-468270-       13796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one dCas9-KRAB-MeCP2 plasmid for cloning of custom sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13796)
  AUTHORS   Borck PC, Boyle I, Jankovic K, Bick N, Foster K, Lau AC, 
            Parker-Burns LI, Lubicki DA, Li T, Borah AA, Lofaso NJ, Das Sharma 
            S, Chan T, Kishen RV, Adeagbo A, Raghavan S, Aquilanti E, Prensner 
            JR, Krill-Burger JM, Golub TR, Campbell CD, Dempster JM, Chan EM, 
            Vazquez F
  TITLE     SKI complex loss renders 9p21.3-deleted or MSI-H cancers dependent 
            on PELO.
  JOURNAL   Nature. 2025 Feb 5. doi: 10.1038/s41586-024-08509-3.
  PUBMED    39910293
REFERENCE   2  (bases 1 to 13796)
  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: "Nature. 
            2025 Feb 5. doi: 10.1038/s41586-024-08509-3."
COMMENT     Sequence Label: pXPR_051_dCas9-KRAB-HA
FEATURES             Location/Qualifiers
     source          1..13796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..1665)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1684..1731)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1779..2108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1847..1866)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2093..2113
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2122..2550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2444..2463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2596..2820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2809..2826
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             2851..3084
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3162..3750
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3268..3279
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3677..3697
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3772..4797)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVHRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(4807..4863)
                     /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             complement(4906..4926)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4933..4953)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4966..4992)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             complement(4972..4998)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(5002..6585,6586..6588,6589..9075,9076..9078,
                     9079..9105))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(5002..9105)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9127..9312)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     protein_bind    complement(9342..9366)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(9392..9603)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        complement(9777..10025)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(9835..9854)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(10005..10025)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    10076..10193
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(10491..10535)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    10720..10953
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    11446..11571
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11618..11798
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(11816..12014)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(11841..11861)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        12016..12395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     12567..12586
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        12661..12765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12766..12834,12835..13626)
                     /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(12984..13003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca cgaagtgcac
     1321 gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc
     1381 ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc
     1441 gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg
     1501 gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc
     1561 ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac
     1621 gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt agttcctcac
     1681 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     1741 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     1801 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     1861 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1921 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1981 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2041 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2101 tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta
     2161 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2221 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2281 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2341 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2401 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2461 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2521 ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc
     2581 tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
     2641 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     2701 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     2761 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     2821 tcgaggctga tcagcgggtt taaacggccc tgctagagat tttccacact gactaaaagg
     2881 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     2941 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     3001 aggctcagat ctggtctaac cagagagacc cagtacaagc aaaaagcaga tcttgtcttc
     3061 gttgggagtg aattagccct tccagtcccc ccttttcttt taaaaagtgg ctaagatcta
     3121 cagctgcctt gtaagtcatt ggtcttaaag tcgacctcga ggcggggagg cggcccaaag
     3181 ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag
     3241 caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt
     3301 cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg atgatttccc
     3361 cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag
     3421 gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag
     3481 gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca
     3541 caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga
     3601 tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat
     3661 taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct
     3721 ttcacaaatt ttgtaatcca gaggttgatt gtcgacttaa cgcgtgttaa ctcattcctt
     3781 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
     3841 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
     3901 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
     3961 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg
     4021 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
     4081 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
     4141 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
     4201 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
     4261 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
     4321 cacatgggga tcagcaatcg cgcatatgaa atcacgccac gtagtgtatt gaccgattcc
     4381 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
     4441 catggcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa
     4501 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctaa attccccaat
     4561 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgatgaa cataacgatc
     4621 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
     4681 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg acacgctgtc
     4741 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatcga
     4801 tccgcccggt ccaggattct cttcgacatc tccggcttgt ttcagcagag agaagtttgt
     4861 tgcgccggat ccgctgctgc cgttgctccc actaccaatg ccgtccactt tccgcttttt
     4921 cttaggatct tccaccttgc gttttttctt gggggagccg ctgcccaggc tggcataatc
     4981 gggcacgtca tagggatagg cgtcgcctcc cagctgagac aggtcgatcc gtgtctcgta
     5041 caggccggtg atgctctggt ggatcagggt ggcgtccagc acctctttgg tgctggtgta
     5101 cctcttccgg tcgatggtgg tgtcaaagta cttgaaggcg gcaggggctc ccagattggt
     5161 cagggtaaac aggtggatga tattctcggc ctgctctctg ataggcttgt ctctgtgctt
     5221 gttgtaggcg ctcagcacct tgtccagatt agcgtcggcc aggatcactc tcttggagaa
     5281 ctcgctgatc tgctcgatga tctcgtccag gtagtgtttg tgctgttcca caaacagctg
     5341 tttctgctca ttatcctcgg gggagccctt cagcttctca tagtggctgg ccaggtacag
     5401 gaagttcaca tatttggagg gcagggccag ttcgtttccc ttctgcagtt cgccggcaga
     5461 ggccagcatt ctcttccggc cgttttccag ctcgaacagg gagtacttag gcagcttgat
     5521 gatcaggtcc tttttcactt ctttgtagcc cttggcttcc agaaagtcga tgggattctt
     5581 ctcgaagctg cttctttcca tgatggtgat ccccagcagc tctttcacac tcttcagttt
     5641 cttggacttg cccttttcca ctttggccac caccagcaca gaataggcca cggtggggct
     5701 gtcgaagccg ccgtacttct tagggtccca gtccttcttt ctggcgatca gcttgtcgct
     5761 gttcctcttg ggcaggatag actctttgct gaagccgcct gtctgcacct cggtcttttt
     5821 cacgatattc acttggggca tagacagcac tttccgcacg gtggcaaagt cccggccctt
     5881 atcccacacg atctcgcctg tttcgccgtt tgtctcgatc agaggccgct tccggatctc
     5941 gccgttggcc agggtaatct cggtcttgaa aaagttcatg atgttgctgt agaagaagta
     6001 cttggcggta gccttgccga tttcctgctc gctcttggcg atcatcttcc gcacgtcgta
     6061 caccttgtag tcgccgtaca cgaactcgct ttccagctta gggtactttt tgatcagggc
     6121 ggttcccacg acggcgttca ggtaggcgtc gtgggcgtgg tggtagttgt tgatctcgcg
     6181 cactttgtaa aactggaaat ccttccggaa atcggacacc agcttggact tcagggtgat
     6241 cactttcact tcccggatca gtttgtcgtt ctcgtcgtac ttagtgttca tccgggagtc
     6301 caggatctgt gccacgtgct ttgtgatctg ccgggtttcc accagctgtc tcttgatgaa
     6361 gccggcctta tccagttcgc tcaggccgcc tctctcggcc ttggtcagat tgtcgaactt
     6421 cctctgggta atcagcttgg cattcagcag ctggcgccag tagttcttca tcttcttcac
     6481 gacctcttcg gagggcacgt tgtcgctctt gccccggttc ttgtcgctcc gagtcagcac
     6541 tttgttatcg atggagtcgt ccttcagaaa gctctgaggc acgatagcgt ccacatcgta
     6601 gtcggacagc cggttgatgt ccagttcctg gtccacgtac atatcccgcc cattctgcag
     6661 gtagtacagg tacagcttct cgttctgcag ctgggtgttt tccacggggt gttctttcag
     6721 gatctggctg cccagctctt tgatgccctc ttcgatccgc ttcattctct cgcggctgtt
     6781 cttctgtccc ttctgggtgg tctggttctc tctggccatt tcgatcacga tgttctcggg
     6841 cttgtgccgg cccatcactt tcacgagctc gtccaccacc ttcactgtct gcaggatgcc
     6901 cttcttaatg gcggggctgc cggccagatt ggcaatgtgc tcgtgcaggc tatcgccctg
     6961 gccggacacc tgggctttct ggatgtcctc tttaaaggtc aggctgtcgt cgtggatcag
     7021 ctgcatgaag tttctgttgg cgaagccgtc ggacttcagg aaatccagga ttgtcttgcc
     7081 ggactgcttg tcccggatgc cgttgatcag cttccggctc agcctgcccc agccggtgta
     7141 tctccgccgc ttcagctgct tcatcacttt gtcgtcgaac aggtgggcat aggttttcag
     7201 ccgttcctcg atcatctctc tgtcctcaaa cagtgtcagg gtcagcacga tatcttccag
     7261 aatgtcctcg ttttcctcat tgtccaggaa gtccttgtcc ttgataattt tcagcagatc
     7321 gtggtatgtg cccagggagg cgttgaaccg atcttccacg ccggagattt ccacggagtc
     7381 gaagcactcg attttcttga agtagtcctc tttcagctgc ttcacggtca ctttccggtt
     7441 ggtcttgaac agcaggtcca cgatggcttt tttctgctcg ccgctcagga aggcgggctt
     7501 tctcattccc tcggtcacgt atttcacttt ggtcagctcg ttgtacacgg tgaagtactc
     7561 gtacagcagg ctgtgcttgg gcagcacctt ctcgttgggc aggttcttat cgaagttggt
     7621 catccgctcg atgaagctct gggcgctggc gcccttgtcc accacttcct cgaagttcca
     7681 gggggtgatg gtttcctcgc tctttctggt catccaggcg aatctgctgt ttcccctggc
     7741 cagagggccc acgtagtagg ggatgcggaa ggtcaggatc ttctcgatct tttcccggtt
     7801 gtccttcagg aatgggtaaa aatcttcctg ccgccgcaga atggcgtgca gctctcccag
     7861 gtggatctgg tgggggatgc tgccgttgtc gaaggtccgc tgcttccgca gcaggtcctc
     7921 tctgttcagc ttcacgagca gttcctcggt gccgtccatc ttttccagga tgggcttgat
     7981 gaacttgtag aactcttcct ggctggctcc gccatcgatg tagccggcgt agccgttctt
     8041 gctctggtcg aagaaaatct ctttgtactt ctcaggcagc tgctgccgca cgagagcttt
     8101 cagcagggtc aggtcctggt ggtgctcgtc gtatctcttg atcatagagg cgctcagggg
     8161 ggccttggtg atctcggtgt tcactctcag gatgtcgctc agcaggatgg cgtcggacag
     8221 gttcttggcg gccagaaaca ggtcggcgta ctggtcgccg atctgggcca gcaggttgtc
     8281 caggtcgtcg tcgtaggtgt ccttgctcag ctgcagtttg gcatcctcgg ccaggtcgaa
     8341 gttgctcttg aagttggggg tcaggcccag gctcagggca atcaggttgc cgaacaggcc
     8401 attcttcttc tcgccgggca gctgggcgat cagattttcc agccgtctgc tcttgctcag
     8461 tctggcagac aggatggcct tggcgtccac gccgctggcg ttgatggggt tttcctcgaa
     8521 cagctggttg taggtctgca ccagctggat gaacagcttg tccacgtcgc tgttgtcggg
     8581 gttcaggtcg ccctcgatca ggaagtggcc ccggaacttg atcatgtggg ccagggccag
     8641 atagatcagc cgcaggtcgg ccttgtcggt gctgtccacc agtttctttc tcaggtggta
     8701 gatggtgggg tacttctcgt ggtaggccac ctcgtccacg atgttgccga agatggggtg
     8761 ccgctcgtgc ttcttatcct cttccaccag gaaggactct tccagtctgt ggaagaagct
     8821 gtcgtccacc ttggccatct cgttgctgaa gatctcttgc agatagcaga tccggttctt
     8881 ccgtctggtg tatcttcttc tggcggttct cttcagccgg gtggcctcgg ctgtttctcc
     8941 gctgtcgaac agcagggcgc cgatcaggtt cttcttgatg ctgtgccggt cggtgttgcc
     9001 cagcaccttg aatttcttgc tgggcacctt gtactcgtcg gtgatcacgg cccagcccac
     9061 agagttggtg ccgatggcca ggccgatgct gtacttcttg tccatcgacc ctccgccgga
     9121 accgccgggc tcttctccct tctccaaccg gaggatcaca tctggcttag taagctgata
     9181 acccaaggaa accaggttct tatagttctc cagcatcaca tttctgtaca cgatctgctg
     9241 agcagtgtcc agcagcttcc actcctccct ggtgaagtcc acaaatacat ccttgaaggt
     9301 caccagtgtc cgggaccagg cagttagtga cttagcatcc atgccaactt ttttgtacaa
     9361 acttgttgat caatcgatgc tagaaccggt cctgtgttct ggcggcaaac ccgttgcgaa
     9421 aaagaacgtt cacggcgact actgcactta tatacggttc tcccccaccc tcgggaaaaa
     9481 ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt ctctaggcac
     9541 cggatcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat gtgcgctctg
     9601 cccactgacg ggcaccggag ccaattccca ctcctttcaa gacctagcta gcgaattcaa
     9661 aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt atttcaactt
     9721 gctacagcat ttctgctgta gctctgaaac agagacgcaa aaaaccggtc gtctccggtg
     9781 tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca agttacggta
     9841 agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat
     9901 tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcta
     9961 attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg
    10021 ccctcggtac cttaattaac caaactggat ctctgctgtc cctgtaataa acccgaaaat
    10081 tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt
    10141 ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga
    10201 tgaatactgc catttgtctg cagaattggc gcacgcagtg ccgatccgtt cactaatcga
    10261 atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt
    10321 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
    10381 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
    10441 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
    10501 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
    10561 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
    10621 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
    10681 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
    10741 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
    10801 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
    10861 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
    10921 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
    10981 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
    11041 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
    11101 agcggccgct tgctgtgcgg tggtcttact tttgttttgc tcttcctcta tcttgtctaa
    11161 agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt
    11221 attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag
    11281 tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga atcgttctag
    11341 ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa
    11401 ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc
    11461 atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc
    11521 ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg
    11581 tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac
    11641 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
    11701 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
    11761 agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa acgcgctgct
    11821 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
    11881 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
    11941 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
    12001 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
    12061 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
    12121 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
    12181 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
    12241 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
    12301 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
    12361 tgattactat taataactag tcaataatca atgtcaacgc gtatatctgg cccgtacatc
    12421 gcgaagcagc gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc
    12481 ttgccttgtt gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc
    12541 agggagcaga tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
    12601 taggggatcg ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg
    12661 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
    12721 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
    12781 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
    12841 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
    12901 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
    12961 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
    13021 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
    13081 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
    13141 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
    13201 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    13261 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    13321 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    13381 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    13441 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    13501 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    13561 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    13621 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    13681 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    13741 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
