LOCUS       sequence-233058-       12303 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Stably express BE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 12303)
  AUTHORS   Li Q, Li Y, Yang S, Huang S, Yan M, Ding Y, Tang W, Lou X, Yin Q, 
            Sun Z, Lu L, Shi H, Wang H, Chen Y, Li J
  TITLE     CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 
            amino acids that are critical for primordial germ cell development.
  JOURNAL   Nat Cell Biol. 2018 Oct 1. pii: 10.1038/s41556-018-0202-4. doi: 
            10.1038/s41556-018-0202-4.
  PUBMED    30275529
REFERENCE   2  (bases 1 to 12303)
  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 Cell 
            Biol. 2018 Oct 1. pii: 10.1038/s41556-018-0202-4. doi: 
            10.1038/s41556-018-0202-4."
COMMENT     Sequence Label: PB-BE3-2NLS
FEATURES             Location/Qualifiers
     source          1..12303
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     15..34
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             67..741
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             67..741
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(197..215)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     465..484
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     659..682
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             745..798
                     /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             799..864
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             871..891
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             913..1599
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(1648..5046,5047..5049,5050..5646,5647,5648..5650,
                     5651..5652,5653..5655,5656..5748)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1648..1671,1672..1674,1675..5748)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5761..6009
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6022..6042
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6051..6068
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6195..6214)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6260..6315
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6314..6333)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6676..6692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6700..6716
                     /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(6724..6730,6731..6748,6749..6754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6769..6790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7491..7525
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7698..7718)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7698..7716)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7737..7753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7761..7777
                     /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(7785..7791,7792..7809,7810..7815))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7830..7851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7968..7985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8139..8727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8219..8238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8898..9689,9690..9758))
                     /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     9521..9540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9759..9863)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9890..10345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9977..9996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10187..10208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10471..10493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10485..10502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10486..10502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10512..10531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10512..10530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   10778..10840
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11924..12303
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
       61 gatccgatgg cctcctccga ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg
      121 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      181 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      241 gacatcctgt cccctcagtt ccagtacggc tccaaggcct acgtgaagca ccccgccgac
      301 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      361 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
      421 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
      481 aagaccatgg gctgggaggc ctccaccgag cggatgtacc ccgaggacgg cgccctgaag
      541 ggcgagatca agatgaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag
      601 accacctaca tggccaagaa gcccgtgcag ctgcccggcg cctacaagac cgacatcaag
      661 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag
      721 ggccgccact ccaccggcgc ccccgagggc agaggaagtc tgctaacatg cggtgacgtc
      781 gaggagaatc ctggcccaga ctataaggac cacgacggag actacaagga tcatgatatt
      841 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
      901 ggagtcacgc gtatgagctc agagactggc ccagtggctg tggaccccac attgagacgg
      961 cggatcgagc cccatgagtt tgaggtattc ttcgatccga gagagctccg caaggagacc
     1021 tgcctgcttt acgaaattaa ttgggggggc cggcactcca tttggcgaca tacatcacag
     1081 aacactaaca agcacgtcga agtcaacttc atcgagaagt tcacgacaga aagatatttc
     1141 tgtccgaaca caaggtgcag cattacctgg tttctcagct ggagcccatg cggcgaatgt
     1201 agtagggcca tcactgaatt cctgtcaagg tatccccacg tcactctgtt tatttacatc
     1261 gcaaggctgt accaccacgc tgacccccgc aatcgacaag gcctgcggga tttgatctct
     1321 tcaggtgtga ctatccaaat tatgactgag caggagtcag gatactgctg gagaaacttt
     1381 gtgaattata gcccgagtaa tgaagcccac tggcctaggt atccccatct gtgggtacga
     1441 ctgtacgttc ttgaactgta ctgcatcata ctgggcctgc ctccttgtct caacattctg
     1501 agaaggaagc agccacagct gacattcttt accatcgctc ttcagtcttg tcattaccag
     1561 cgactgcccc cacacattct ctgggccacc gggttgaaaa gcggcagcga gactcccggg
     1621 acctcagagt ccgccacacc cgaaagtgat aaaaagtatt ctattggttt agccatcggc
     1681 actaattccg ttggatgggc tgtcataacc gatgaataca aagtaccttc aaagaaattt
     1741 aaggtgttgg ggaacacaga ccgtcattcg attaaaaaga atcttatcgg tgccctccta
     1801 ttcgatagtg gcgaaacggc agaggcgact cgcctgaaac gaaccgctcg gagaaggtat
     1861 acacgtcgca agaaccgaat atgttactta caagaaattt ttagcaatga gatggccaaa
     1921 gttgacgatt ctttctttca ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa
     1981 catgaacggc accccatctt tggaaacata gtagatgagg tggcatatca tgaaaagtac
     2041 ccaacgattt atcacctcag aaaaaagcta gttgactcaa ctgataaagc ggacctgagg
     2101 ttaatctact tggctcttgc ccatatgata aagttccgtg ggcactttct cattgagggt
     2161 gatctaaatc cggacaactc ggatgtcgac aaactgttca tccagttagt acaaacctat
     2221 aatcagttgt ttgaagagaa ccctataaat gcaagtggcg tggatgcgaa ggctattctt
     2281 agcgcccgcc tctctaaatc ccgacggcta gaaaacctga tcgcacaatt acccggagag
     2341 aagaaaaatg ggttgttcgg taaccttata gcgctctcac taggcctgac accaaatttt
     2401 aagtcgaact tcgacttagc tgaagatgcc aaattgcagc ttagtaagga cacgtacgat
     2461 gacgatctcg acaatctact ggcacaaatt ggagatcagt atgcggactt atttttggct
     2521 gccaaaaacc ttagcgatgc aatcctccta tctgacatac tgagagttaa tactgagatt
     2581 accaaggcgc cgttatccgc ttcaatgatc aaaaggtacg atgaacatca ccaagacttg
     2641 acacttctca aggccctagt ccgtcagcaa ctgcctgaga aatataagga aatattcttt
     2701 gatcagtcga aaaacgggta cgcaggttat attgacggcg gagcgagtca agaggaattc
     2761 tacaagttta tcaaacccat attagagaag atggatggga cggaagagtt gcttgtaaaa
     2821 ctcaatcgcg aagatctact gcgaaagcag cggactttcg acaacggtag cattccacat
     2881 caaatccact taggcgaatt gcatgctata cttagaaggc aggaggattt ttatccgttc
     2941 ctcaaagaca atcgtgaaaa gattgagaaa atcctaacct ttcgcatacc ttactatgtg
     3001 ggacccctgg cccgagggaa ctctcggttc gcatggatga caagaaagtc cgaagaaacg
     3061 attactccat ggaattttga ggaagttgtc gataaaggtg cgtcagctca atcgttcatc
     3121 gagaggatga ccaactttga caagaattta ccgaacgaaa aagtattgcc taagcacagt
     3181 ttactttacg agtatttcac agtgtacaat gaactcacga aagttaagta tgtcactgag
     3241 ggcatgcgta aacccgcctt tctaagcgga gaacagaaga aagcaatagt agatctgtta
     3301 ttcaagacca accgcaaagt gacagttaag caattgaaag aggactactt taagaaaatt
     3361 gaatgcttcg attctgtcga gatctccggg gtagaagatc gatttaatgc gtcacttggt
     3421 acgtatcatg acctcctaaa gataattaaa gataaggact tcctggataa cgaagagaat
     3481 gaagatatct tagaagatat agtgttgact cttaccctct ttgaagatcg ggaaatgatt
     3541 gaggaaagac taaaaacata cgctcacctg ttcgacgata aggttatgaa acagttaaag
     3601 aggcgtcgct atacgggctg gggacgattg tcgcggaaac ttatcaacgg gataagagac
     3661 aagcaaagtg gtaaaactat tctcgatttt ctaaagagcg acggcttcgc caataggaac
     3721 tttatgcagc tgatccatga tgactcttta accttcaaag aggatataca aaaggcacag
     3781 gtttccggac aaggggactc attgcacgaa catattgcga atcttgctgg ttcgccagcc
     3841 atcaaaaagg gcatactcca gacagtcaaa gtagtggatg agctagttaa ggtcatggga
     3901 cgtcacaaac cggaaaacat tgtaatcgag atggcacgcg aaaatcaaac gactcagaag
     3961 gggcaaaaaa acagtcgaga gcggatgaag agaatagaag agggtattaa agaactgggc
     4021 agccagatct taaaggagca tcctgtggaa aatacccaat tgcagaacga gaaactttac
     4081 ctctattacc tacaaaatgg aagggacatg tatgttgatc aggaactgga cataaaccgt
     4141 ttatctgatt acgacgtcga tcacattgta ccccaatcct ttttgaagga cgattcaatc
     4201 gacaataaag tgcttacacg ctcggataag aaccgaggga aaagtgacaa tgttccaagc
     4261 gaggaagtcg taaagaaaat gaagaactat tggcggcagc tcctaaatgc gaaactgata
     4321 acgcaaagaa agttcgataa cttaactaaa gctgagaggg gtggcttgtc tgaacttgac
     4381 aaggccggat ttattaaacg tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca
     4441 cagatactag attcccgaat gaatacgaaa tacgacgaga acgataagct gattcgggaa
     4501 gtcaaagtaa tcactttaaa gtcaaaattg gtgtcggact tcagaaagga ttttcaattc
     4561 tataaagtta gggagataaa taactaccac catgcgcacg acgcttatct taatgccgtc
     4621 gtagggaccg cactcattaa gaaatacccg aagctagaaa gtgagtttgt gtatggtgat
     4681 tacaaagttt atgacgtccg taagatgatc gcgaaaagcg aacaggagat aggcaaggct
     4741 acagccaaat acttctttta ttctaacatt atgaatttct ttaagacgga aatcactctg
     4801 gcaaacggag agatacgcaa acgaccttta attgaaacca atggggagac aggtgaaatc
     4861 gtatgggata agggccggga cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc
     4921 aacatagtaa agaaaactga ggtgcagacc ggagggtttt caaaggaatc gattcttcca
     4981 aaaaggaata gtgataagct catcgctcgt aaaaaggact gggacccgaa aaagtacggt
     5041 ggcttcgata gccctacagt tgcctattct gtcctagtag tggcaaaagt tgagaaggga
     5101 aaatccaaga aactgaagtc agtcaaagaa ttattgggga taacgattat ggagcgctcg
     5161 tcttttgaaa agaaccccat cgacttcctt gaggcgaaag gttacaagga agtaaaaaag
     5221 gatctcataa ttaaactacc aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg
     5281 atgttggcta gcgccggaga gcttcaaaag gggaacgaac tcgcactacc gtctaaatac
     5341 gtgaatttcc tgtatttagc gtcccattac gagaagttga aaggttcacc tgaagataac
     5401 gaacagaagc aactttttgt tgagcagcac aaacattatc tcgacgaaat catagagcaa
     5461 atttcggaat tcagtaagag agtcatccta gctgatgcca atctggacaa agtattaagc
     5521 gcatacaaca agcacaggga taaacccata cgtgagcagg cggaaaatat tatccatttg
     5581 tttactctta ccaacctcgg cgctccagcc gcattcaagt attttgacac aacgatagat
     5641 cgcaaacgat acacttctac caaggaggtg ctagacgcga cactgattca ccaatccatc
     5701 acgggattat atgaaactcg gatagatttg tcacagcttg ggggtgactc tggtggttct
     5761 actaatctgt cagatattat tgaaaaggag accggtaagc aactggttat ccaggaatcc
     5821 atcctcatgc tcccagagga ggtggaagaa gtcattggga acaagccgga aagcgatata
     5881 ctcgtgcaca ccgcctacga cgagagcacc gacgagaatg tcatgcttct gactagcgac
     5941 gcccctgaat acaagccttg ggctctggtc atacaggata gcaacggtga gaacaagatt
     6001 aagatgctct ctggtggttc tcccaagaag aagaggaaag tctaaccggt catcatcacc
     6061 atcaccattg agtttaaacc cgctgatcag cacgcgtgct agcttcgaag ctaattcact
     6121 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     6181 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     6241 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     6301 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     6361 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     6421 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     6481 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     6541 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     6601 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     6661 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6721 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6781 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6841 gcggatcgat ctgacaatgt tcagtgcaga gactcggcta cgcctcgtgg actttgaagt
     6901 tgaccaacaa tgtttattct tacctctaat agtcctctgt ggcaaggtca agattctgtt
     6961 agaagccaat gaagaacctg gttgttcaat aacattttgt tcgtctaata tttcactacc
     7021 gcttgacgtt ggctgcactt catgtacctc atctataaac gcttcttctg tatcgctctg
     7081 gacgtcatct tcacttacgt gatctgatat ttcactgtca gaatcctcac caacaagctc
     7141 gtcatcgctt tgcagaagag cagagaggat atgctcatcg tctaaagaac tacccatttt
     7201 attatatatt agtcacgata tctataacaa gaaaatatat atataataag ttatcacgta
     7261 agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa
     7321 agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta
     7381 ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca
     7441 cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa
     7501 ttttacgcag actatctttc tagggttaaa aaagatttgc gctttactcg acctaaactt
     7561 taaacacgtc atagaatctt cgtttgacaa aaaccacatt gtggccaagc tgtgtgacgc
     7621 gacgcgcgct aaagaatggc aaaccaagtc gcgcgagcgt cgacctcgag ggggggcccg
     7681 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     7741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7801 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     8041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     8101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     8161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     8221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     8341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     8581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8941 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     9001 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9061 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     9901 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     9961 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
    10021 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
    10081 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    10141 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
    10201 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
    10261 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
    10321 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
    10381 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
    10441 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
    10501 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggccgcccgg
    10561 tttatcgtta atatggatca atttgaacag ttgattaacg tgtctctgct caagtctttg
    10621 atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca agtcgatgag cgaaaaacta
    10681 aaaaggctag aatacgacaa tctcacagac agcgttgaga tatacggtat tcacgacagc
    10741 aggctgaata ataaaaaaat tagaaactat tatttaaccc tagaaagata atcatattgt
    10801 gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg gtctgtatat
    10861 cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca cttacatact
    10921 aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa aaaacaaaaa
    10981 ctcaaaattt cttctataaa gtaacaaaac ttttaaacat tctctctttt acaaaaataa
    11041 acttattttg tactttaaaa acagtcatgt tgtattataa aataagtaat tagcttaact
    11101 tatacataat agaaacaaat tatacttatt agtcagtcag aaacaacttt ggcacatatc
    11161 aatattatgc tctcgacaaa taactttttt gcattttttg cacgatgcat ttgcctttcg
    11221 ccttatttta gaggggcagt aagtacagta agtacgtttt ttcattactg gctcttcagt
    11281 actgtcatct gatgtaccag gcacttcatt tggcaaaata ttagagatat tatcgcgcaa
    11341 atatctcttc aaagtaggag cttctaaacg cttacgcata aacgatgacg tcaggctcat
    11401 gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt tctcccttgc tactgacatt
    11461 atggctgtat ataataaaag aatttatgca ggcaatgttt atcattccgt acaataatgc
    11521 cataggccac ctattcgtct tcctactgca ggtcatcaca gaacacattt ggtctagcgt
    11581 gtccactccg cctttagttt gattataata cataaccatt tgcggtttac cggtactttc
    11641 gttgatagaa gcatcctcat cacaagatga taataagtat accatcttag ctggcttcgg
    11701 tttatatgag acgagagtaa ggggtccgtc aaaacaaaac atcgatgttc ccactggcct
    11761 ggagcgactg tttttcagta cttccggtat ctcgcgtttg tttgatcgca cggttcccac
    11821 aatggttaat tcgagctcgc ccaaaccggg cgcgccgggc atttaaatgg aattcgatgc
    11881 cctatatatg gatccaattg caatgatcat catgacagat ctcgacattg attattgact
    11941 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    12001 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    12061 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    12121 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    12181 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    12241 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
    12301 atg
//
