LOCUS       sequence-273028-       13013 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express guide RNA 1 for mouse Myc.
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 Apr 14, 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-sgMyc_2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    184..301
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        352..600
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     352..372
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     523..542
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        622..697
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        759..970
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      989..998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             995..5098
                     /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             995..5098
                     /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             5099..5146
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5147..5170
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5180..5236
                     /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             5237..5833
                     /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     5730..5750
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5849..6437
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5902..5922)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6320..6331)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6509..6742
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6768..6785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6774..6998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7044..7472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7131..7150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7341..7362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7481..7501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7486..7815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7666..7801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7728..7747
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7863..7910
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7929..8303
                     /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    8433..8554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8470..8489)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8524..8543
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8603..8619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8603..8619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8616..8638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8627..8643
                     /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(8651..8657,8658..8675,8676..8681))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8696..8717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8834..8851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9005..9593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9085..9104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9764..10555,10556..10624))
                     /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     10387..10406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10625..10729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10804..10823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10995..11374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11376..11574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11529..11549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11592..11772
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11819..11944
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12437..12670
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12855..12899
                     /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 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
       61 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
      121 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
      181 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
      241 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
      301 tcgggtttat tacagggaca gcagagatcc agtttggtta attaaggtac cgagggccta
      361 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga
      421 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
      481 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
      541 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
      601 gagaggcaaa cccctgccaa ggttttagag ctagaaatag caagttaaaa taaggctagt
      661 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgaattcg ctagctaggt
      721 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
      781 agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg
      841 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
      901 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
      961 cagaacacag gaccggttct agagcgctgc caccatggac aagaagtaca gcatcggcct
     1021 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     1081 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     1141 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     1201 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1261 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1321 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1381 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     1441 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     1501 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1561 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     1621 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     1681 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     1741 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     1801 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     1861 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     1921 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     1981 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     2041 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     2101 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     2161 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     2221 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2281 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2341 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     2401 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     2461 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     2521 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     2581 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     2641 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     2701 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     2761 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     2821 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     2881 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     2941 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     3001 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     3061 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     3121 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     3181 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3241 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     3301 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     3361 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     3421 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     3481 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     3541 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     3601 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     3661 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     3721 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     3781 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     3841 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     3901 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     3961 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     4021 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     4081 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     4141 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     4201 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4261 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     4321 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     4381 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     4441 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     4501 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4561 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     4621 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     4681 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     4741 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     4801 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     4861 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     4921 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     4981 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     5041 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     5101 gcgacctgcc gccacaaaga aggctggaca ggctaagaag aagaaagatt acaaagacga
     5161 tgacgataag ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga
     5221 agagaatcct ggaccgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     5281 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5341 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5401 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5461 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5521 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5581 ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg
     5641 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5701 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5761 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     5821 gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5881 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5941 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6001 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6061 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6121 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6181 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6241 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6301 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6361 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6421 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     6481 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     6541 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6601 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6661 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6721 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     6781 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6841 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     6901 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     6961 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     7021 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7081 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7141 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7201 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     7261 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7321 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7381 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     7441 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     7501 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7561 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     7621 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     7681 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     7741 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     7801 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     7861 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     7921 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     7981 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     8041 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     8101 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     8161 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     8221 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     8281 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     8341 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     8401 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     8461 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     8521 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     8581 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     8641 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     8701 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8761 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8821 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8881 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8941 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     9001 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     9061 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     9121 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     9181 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     9241 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     9301 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     9361 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     9421 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     9481 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     9541 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     9601 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9661 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9721 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     9841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    10021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    10081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    10141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    10201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    10261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    10321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    10381 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    10441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    10501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    10561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    10621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    10681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10741 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
    10801 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
    10861 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
    10921 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
    10981 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
    11041 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
    11101 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
    11161 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
    11221 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
    11281 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
    11341 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
    11401 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
    11461 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
    11521 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
    11581 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
    11641 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
    11701 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
    11761 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
    11821 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
    11881 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
    11941 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
    12001 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
    12061 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
    12121 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
    12181 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
    12241 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
    12301 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
    12361 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
    12421 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
    12481 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
    12541 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
    12601 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
    12661 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
    12721 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
    12781 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
    12841 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
    12901 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
    12961 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat aga
//
