LOCUS       sequence-336078-       12823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid used for Crispr/cas9 based disruption of human DNMT1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12823)
  AUTHORS   Jonsson ME, Ludvik Brattas P, Gustafsson C, Petri R, Yudovich D, 
            Pircs K, Verschuere S, Madsen S, Hansson J, Larsson J, Mansson R, 
            Meissner A, Jakobsson J
  TITLE     Activation of neuronal genes via LINE-1 elements upon global DNA 
            demethylation in human neural progenitors.
  JOURNAL   Nat Commun. 2019 Jul 18;10(1):3182. doi: 10.1038/s41467-019-11150-8.
  PUBMED    31320637
REFERENCE   2  (bases 1 to 12823)
  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: "Nat 
            Commun."; date: "2019-07-18"; volume: "10(1):3182. doi"; pages: " 
            10.1038/s41467-019-11150-8"
COMMENT     Sequence Label: pLV.U6.gDNMT1.Cas9-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..12823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2878..2953
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3015..3226
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     3346..3368
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3430..3449
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3483..3492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3489..7592
                     /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             3489..7592
                     /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             3489..7592
                     /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             7605..7625
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7641..7694
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(7695..7697,7698..7700,7701..8414)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7695..7697,7698..7700,7701..8411)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7740..7761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8001..8020)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8348..8369
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             8575..8808
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8834..8851)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8840..9064
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9110..9538
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9197..9216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9407..9428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9547..9567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9552..9881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9732..9867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9794..9813
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9929..9976
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9995..10369
                     /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    10499..10620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10536..10555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10590..10609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10669..10685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10669..10685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10682..10704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10693..10709
                     /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(10717..10723,10724..10741,10742..10747))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10762..10783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10900..10917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11071..11659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11151..11170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11830..12621,12622..12690))
                     /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     12453..12472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12691..12795)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggat gtgggaccct gcggcccgtt
     2881 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     2941 accgagtcgg tgcttttttg aattcgctag ctaggtcttg aaaggagtgg gaattggctc
     3001 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     3061 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     3121 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     3181 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggtatgggcg
     3241 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     3301 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     3361 gtgtgaccgg cggctctagc gcctctgcta accatgttca tgccttcttc tttttcctac
     3421 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcgcgg
     3481 ccgccaccat ggacaagaag tactccattg ggctcgatat cggcacaaac agcgtcggct
     3541 gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata
     3601 ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac tccggggaga
     3661 cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc agaaagaatc
     3721 ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat gactctttct
     3781 tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag cgccacccaa
     3841 tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc atatatcatc
     3901 tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc tatctcgcgc
     3961 tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg aacccagaca
     4021 acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag cttttcgaag
     4081 agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct aggctgtcca
     4141 aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag aacggcctgt
     4201 ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct aacttcgacc
     4261 tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat ctcgacaatc
     4321 tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag aacctgtcag
     4381 acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa gctccgctga
     4441 gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg ctgaaggccc
     4501 ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag tctaaaaatg
     4561 gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa tttattaagc
     4621 ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac agagaagatc
     4681 tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt cacctgggcg
     4741 aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa gataacaggg
     4801 aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc ctcgcccggg
     4861 gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact ccctggaact
     4921 tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg atgactaact
     4981 ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact
     5041 tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg agaaagccag
     5101 cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag acgaaccgga
     5161 aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt ttcgactctg
     5221 ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat cacgatctcc
     5281 tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac attcttgagg
     5341 acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa cgcttgaaaa
     5401 cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc cgatatacag
     5461 gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag agtggaaaga
     5521 caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg cagttgatcc
     5581 atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct ggccaggggg
     5641 acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa aagggaatac
     5701 tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat aagcccgaga
     5761 atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag aagaacagta
     5821 gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa atccttaagg
     5881 aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac tacctgcaga
     5941 acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc gactacgacg
     6001 tggatcatat cgtgccccag tcttttctca aagatgattc tattgataat aaagtgttga
     6061 caagatccga taaaaataga gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga
     6121 aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa cggaagttcg
     6181 ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca
     6241 aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt ctcgattcac
     6301 gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa gttattactc
     6361 tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag gtgagagaga
     6421 tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc actgcactta
     6481 tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa gtgtacgatg
     6541 ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct aagtacttct
     6601 tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat ggagagattc
     6661 ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg gacaagggta
     6721 gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga
     6781 ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg aacagcgaca
     6841 agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc gattctccta
     6901 cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct aaaaaactca
     6961 aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc
     7021 ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc atcattaagc
     7081 ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc gctagtgcgg
     7141 gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat ttcttgtatc
     7201 tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag aagcagctgt
     7261 tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc gaattctcca
     7321 aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac aataagcaca
     7381 gggataagcc catcagggag caggcagaaa acattatcca cttgtttact ctgaccaact
     7441 tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag cggtacacct
     7501 ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg ctctatgaaa
     7561 caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag aagaagagga
     7621 aggtgaggtc cggcggcgga gagggcagag gaagtcttct aacatgcggt gacgtggagg
     7681 agaatcccgg cccaatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     7741 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     7801 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     7861 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     7921 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     7981 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     8041 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     8101 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     8161 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     8221 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     8281 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     8341 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     8401 agctgtacaa gtaaccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     8461 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     8521 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     8581 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     8641 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     8701 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     8761 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
     8821 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     8881 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     8941 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     9001 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     9061 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     9121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     9181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     9241 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     9301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     9361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     9421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     9481 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     9541 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     9601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     9661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     9721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     9781 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     9841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     9901 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     9961 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
    10021 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
    10081 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
    10141 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
    10201 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
    10261 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
    10321 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
    10381 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
    10441 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
    10501 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
    10561 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
    10621 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
    10681 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    10741 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    10801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    10861 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    10921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    10981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    11041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    11101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    11161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    11221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    11281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    11341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    11401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
    11461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
    11521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    11581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    11641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    11701 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
    11761 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    11821 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    11881 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    11941 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    12001 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    12061 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    12121 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    12181 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    12241 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    12301 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    12361 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    12421 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    12481 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    12541 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    12601 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    12661 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    12721 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    12781 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
