LOCUS       sequence-273024-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express guide RNA 2 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 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-sgMyc_1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    139..256
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        307..555
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     307..327
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     478..497
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        577..652
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        714..925
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      944..953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             950..5053
                     /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             950..5053
                     /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             5054..5101
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5102..5125
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5135..5191
                     /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             5192..5788
                     /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     5685..5705
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5804..6392
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5857..5877)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6275..6286)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6464..6697
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6723..6740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6729..6953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6999..7427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7086..7105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7296..7317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7436..7456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7441..7770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7621..7756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7683..7702
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7818..7865
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7884..8258
                     /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    8388..8509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8425..8444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8479..8498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8558..8574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8558..8574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8571..8593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8582..8598
                     /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(8606..8612,8613..8630,8631..8636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8651..8672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8789..8806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8960..9548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9040..9059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9719..10510,10511..10579))
                     /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     10342..10361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10580..10684)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10759..10778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10950..11329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11331..11529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11484..11504
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11547..11727
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11774..11899
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12392..12625
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12810..12854
                     /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 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
       61 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
      121 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      181 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      241 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa
      301 ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
      361 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      421 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      481 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      541 aaaggacgaa acaccggtca atgcactcgg acgcgggttt tagagctaga aatagcaagt
      601 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
      661 attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
      721 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
      781 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      841 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      901 caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa
      961 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     1021 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     1081 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     1141 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     1201 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     1261 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     1321 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     1381 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     1441 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     1501 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     1561 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     1621 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     1681 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     1741 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     1801 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     1861 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     1921 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     1981 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     2041 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     2101 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     2161 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     2221 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     2281 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     2341 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     2401 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     2461 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     2521 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     2581 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     2641 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     2701 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     2761 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     2821 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     2881 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     2941 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     3001 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     3061 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     3121 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     3181 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     3241 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     3301 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     3361 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     3421 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     3481 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     3541 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     3601 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     3661 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     3721 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     3781 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     3841 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     3901 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     3961 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     4021 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     4081 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     4141 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     4201 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     4261 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     4321 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     4381 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     4441 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     4501 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     4561 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     4621 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     4681 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     4741 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     4801 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     4861 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     4921 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     4981 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     5041 gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
     5101 agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca
     5161 agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc
     5221 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5281 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5341 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5401 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5461 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5521 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
     5581 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5641 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5701 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5761 gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta
     5821 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5881 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5941 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6001 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6061 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6121 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6181 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     6241 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6301 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6361 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     6421 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     6481 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6541 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6601 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6661 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
     6721 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6781 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6841 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     6901 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     6961 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     7021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     7081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     7441 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     7501 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     7561 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     7621 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     7681 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     7741 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     7801 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     7861 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     7921 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     7981 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     8041 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     8101 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     8161 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     8221 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
     8281 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     8341 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     8401 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     8461 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     8521 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     8581 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     8641 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     8701 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     8761 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     8821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     8881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     8941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     9001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     9061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     9121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     9181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     9241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     9301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     9361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     9421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     9481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     9541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     9601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     9661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     9721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     9781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     9841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     9901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     9961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    10021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    10081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    10141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    10201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    10261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    10321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    10381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    10441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    10501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    10561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    10621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    10681 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
    10741 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
    10801 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
    10861 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
    10921 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
    10981 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
    11041 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
    11101 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
    11161 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
    11221 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
    11281 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
    11341 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
    11401 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
    11461 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
    11521 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
    11581 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
    11641 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
    11701 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
    11761 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
    11821 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
    11881 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
    11941 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
    12001 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
    12061 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
    12121 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
    12181 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
    12241 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
    12301 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
    12361 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
    12421 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
    12481 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
    12541 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
    12601 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
    12661 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
    12721 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
    12781 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
    12841 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
    12901 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
    12961 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc cca
//
