LOCUS       sequence-273745-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express guide RNA 1 for 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_1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    236..469
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             654..698
                     /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    996..1113
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1164..1412
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1164..1184
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1335..1354
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1434..1509
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1571..1782
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1801..1810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1807..5910
                     /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             1807..5910
                     /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             5911..5958
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5959..5982
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5992..6048
                     /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             6049..6645
                     /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     6542..6562
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6661..7249
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6714..6734)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7132..7143)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7321..7554
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7580..7597)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7586..7810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7856..8284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7943..7962)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8153..8174
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8293..8313)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8298..8627
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8478..8613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8540..8559
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8675..8722
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8741..9115
                     /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    9245..9366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9282..9301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9336..9355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9415..9431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9415..9431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9428..9450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9439..9455
                     /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(9463..9469,9470..9487,9488..9493))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9508..9529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9646..9663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9817..10405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9897..9916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10576..11367,11368..11436))
                     /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     11199..11218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11437..11541)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11616..11635)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11807..12186
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12188..12386
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     12341..12361
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             12404..12584
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12631..12756
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
       61 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
      121 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
      181 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
      241 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
      301 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
      361 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
      421 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
      481 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
      541 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
      601 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
      661 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
      721 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
      781 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
      841 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
      901 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
      961 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     1021 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     1081 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     1141 ccagtttggt taattaaggt accgagggcc tatttcccat gattccttca tatttgcata
     1201 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     1261 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     1321 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     1381 ctttatatat cttgtggaaa ggacgaaaca ccgtaatgtg aaccggttca caggttttag
     1441 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1501 agtcggtgct tttttgaatt cgctagctag gtcttgaaag gagtgggaat tggctccggt
     1561 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     1621 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     1681 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     1741 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggaccggtt ctagagcgct
     1801 gccaccatgg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     1861 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     1921 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     1981 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     2041 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     2101 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     2161 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     2221 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     2281 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     2341 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     2401 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     2461 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     2521 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     2581 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     2641 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     2701 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     2761 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     2821 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     2881 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     2941 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     3001 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     3061 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     3121 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     3181 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     3241 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     3301 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     3361 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     3421 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     3481 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     3541 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     3601 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     3661 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     3721 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     3781 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     3841 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     3901 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     3961 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     4021 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     4081 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     4141 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     4201 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     4261 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     4321 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     4381 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     4441 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     4501 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     4561 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     4621 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     4681 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     4741 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     4801 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     4861 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     4921 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     4981 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     5041 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     5101 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     5161 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     5221 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     5281 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     5341 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     5401 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     5461 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     5521 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     5581 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     5641 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     5701 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     5761 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     5821 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     5881 cggatcgacc tgtctcagct gggaggcgac aagcgacctg ccgccacaaa gaaggctgga
     5941 caggctaaga agaagaaaga ttacaaagac gatgacgata agggatccgg cgcaacaaac
     6001 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgac cgagtacaag
     6061 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     6121 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     6181 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     6241 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     6301 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     6361 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     6421 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     6481 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     6541 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     6601 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
     6661 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     6721 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     6781 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     6841 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     6901 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     6961 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     7021 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     7081 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     7141 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     7201 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     7261 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     7321 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     7381 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7441 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7501 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
     7561 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     7621 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     7681 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     7741 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     7801 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     7861 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     7921 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     7981 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     8041 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     8101 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     8161 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     8221 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     8281 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     8341 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     8401 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     8461 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     8521 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     8581 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     8641 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     8701 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
     8761 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
     8821 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     8881 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     8941 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
     9001 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
     9061 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
     9121 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     9181 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     9241 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9361 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     9421 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     9481 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     9541 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     9601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     9661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     9841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     9901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     9961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    10381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    10441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    10501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    10561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    10621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    10681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    10741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    10801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    10861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    10921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    10981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    11041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    11101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    11161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    11221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    11281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    11341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    11401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    11461 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    11521 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    11581 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
    11641 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
    11701 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
    11761 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
    11821 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
    11881 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
    11941 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
    12001 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
    12061 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
    12121 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
    12181 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
    12241 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
    12301 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
    12361 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
    12421 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
    12481 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
    12541 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
    12601 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
    12661 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
    12721 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
    12781 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
    12841 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
    12901 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
    12961 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aag
//
