LOCUS       sequence-273026-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express guide RNA 2 for mouse Dnmt1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Fu X, Wu X, Djekidel MN, Zhang Y
  TITLE     Myc and Dnmt1 impede the pluripotent to totipotent state transition 
            in embryonic stem cells.
  JOURNAL   Nat Cell Biol. 2019 Jul;21(7):835-844. doi: 
            10.1038/s41556-019-0343-0. Epub 2019 Jun 17.
  PUBMED    31209294
REFERENCE   2  (bases 1 to 13013)
  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"; doi: 
            "10.1038/s41556-019-0343-0"; journalName: "Nat Cell Biol"; date: 
            "2019-07"; volume: "21"; issue: "7"; pages: "835-844"
COMMENT     Sequence Label: lentiCRISPR v2-sgDnmt1_2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    203..320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        371..619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     371..391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     542..561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        641..716
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        778..989
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1008..1017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1014..5117
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1014..5117
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5118..5165
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5166..5189
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5199..5255
                     /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             5256..5852
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5749..5769
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5868..6456
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5921..5941)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6339..6350)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6528..6761
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6787..6804)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6793..7017
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7063..7491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7150..7169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7360..7381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7500..7520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7505..7834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7685..7820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7747..7766
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7882..7929
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7948..8322
                     /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"
     polyA_signal    8452..8573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8489..8508)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8543..8562
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8622..8638)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8622..8638)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8635..8657)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8646..8662
                     /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)."
     promoter        complement(join(8670..8676,8677..8694,8695..8700))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8715..8736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8853..8870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9024..9612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9104..9123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9783..10574,10575..10643))
                     /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     10406..10425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10644..10748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10823..10842)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11014..11393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11395..11593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11548..11568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11611..11791
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11838..11963
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12456..12689
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12874..12918
                     /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"
ORIGIN
        1 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
       61 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
      121 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
      181 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
      241 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
      301 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
      361 ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
      421 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
      481 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
      541 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
      601 gtggaaagga cgaaacaccg catcacggct cacttcacga gttttagagc tagaaatagc
      661 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      721 ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
      781 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
      841 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
      901 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
      961 ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc accatggaca
     1021 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
     1081 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     1141 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     1201 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     1261 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     1321 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     1381 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     1441 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     1501 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     1561 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     1621 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     1681 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     1741 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     1801 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     1861 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     1921 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     1981 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     2041 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     2101 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     2161 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     2221 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     2281 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     2341 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     2401 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     2461 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     2521 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     2581 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     2641 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     2701 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     2761 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     2821 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     2881 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     2941 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     3001 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     3061 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     3121 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     3181 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     3241 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     3301 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     3361 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     3421 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     3481 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     3541 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     3601 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     3661 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     3721 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     3781 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     3841 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     3901 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     3961 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     4021 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     4081 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     4141 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     4201 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     4261 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     4321 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     4381 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     4441 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     4501 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     4561 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     4621 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     4681 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     4741 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     4801 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     4861 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     4921 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     4981 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     5041 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     5101 ctcagctggg aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga
     5161 agaaagatta caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga
     5221 aacaagccgg agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc
     5281 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     5341 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     5401 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5461 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5521 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5581 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag
     5641 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5701 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5761 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5821 cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg
     5881 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5941 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     6001 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6061 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6121 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6181 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6241 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     6301 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6361 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6421 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta
     6481 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     6541 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     6601 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6661 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6721 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct
     6781 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     6841 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     6901 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     6961 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     7021 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     7081 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7141 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7201 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7261 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7321 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7381 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     7441 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     7501 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     7561 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     7621 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     7681 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     7741 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     7801 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     7861 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     7921 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     7981 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     8041 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     8101 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     8161 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     8221 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     8281 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     8341 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     8401 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     8461 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     8521 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     8581 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     8641 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     8701 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     8761 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     8821 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     8881 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     8941 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     9001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     9061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     9121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     9181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     9241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     9301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     9361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     9421 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     9481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     9541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     9601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     9661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     9721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     9781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     9841 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     9901 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     9961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    10021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    10081 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    10141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    10201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    10261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    10321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    10381 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    10441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    10501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    10561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    10621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    10681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    10741 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
    10801 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
    10861 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
    10921 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
    10981 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
    11041 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
    11101 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
    11161 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
    11221 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
    11281 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
    11341 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
    11401 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
    11461 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
    11521 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
    11581 tctatataag cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc
    11641 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
    11701 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    11761 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
    11821 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
    11881 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
    11941 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
    12001 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
    12061 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
    12121 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
    12181 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
    12241 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
    12301 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
    12361 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
    12421 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
    12481 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
    12541 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
    12601 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
    12661 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
    12721 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
    12781 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
    12841 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
    12901 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
    12961 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgt
//
