LOCUS       sequence-233754-       12276 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 12276)
  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 12276)
  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-BE3-3NLS
FEATURES             Location/Qualifiers
     source          1..12276
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     8..27
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             60..734
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             60..734
                     /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(190..208)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     458..477
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     652..675
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             738..791
                     /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             792..857
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             864..884
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             906..1592
                     /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(1641..5039,5040..5042,5043..5639,5640,5641..5643,
                     5644..5645,5646..5648,5649..5741)
                     /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(1641..1664,1665..1667,1668..5741)
                     /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             5754..6002
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6015..6035
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6042..6062
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6168..6187)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6233..6288
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6287..6306)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6649..6665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6673..6689
                     /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(6697..6703,6704..6721,6722..6727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6742..6763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7464..7498
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7671..7691)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7671..7689)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7710..7726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7734..7750
                     /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(7758..7764,7765..7782,7783..7788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7803..7824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7941..7958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8112..8700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8192..8211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8871..9662,9663..9731))
                     /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     9494..9513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9732..9836)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9863..10318
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9950..9969)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10160..10181
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10444..10466
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10458..10475
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10459..10475
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10485..10504
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10485..10503
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   10751..10813
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11897..12276
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgatccga
       61 tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct
      121 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      181 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
      241 tgtcccctca gttccagtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      301 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      361 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
      421 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
      481 tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga
      541 tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct
      601 acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca
      661 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc
      721 actccaccgg cgcccccgag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga
      781 atcctggccc agactataag gaccacgacg gagactacaa ggatcatgat attgattaca
      841 aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtca
      901 cgcgtatgag ctcagagact ggcccagtgg ctgtggaccc cacattgaga cggcggatcg
      961 agccccatga gtttgaggta ttcttcgatc cgagagagct ccgcaaggag acctgcctgc
     1021 tttacgaaat taattggggg ggccggcact ccatttggcg acatacatca cagaacacta
     1081 acaagcacgt cgaagtcaac ttcatcgaga agttcacgac agaaagatat ttctgtccga
     1141 acacaaggtg cagcattacc tggtttctca gctggagccc atgcggcgaa tgtagtaggg
     1201 ccatcactga attcctgtca aggtatcccc acgtcactct gtttatttac atcgcaaggc
     1261 tgtaccacca cgctgacccc cgcaatcgac aaggcctgcg ggatttgatc tcttcaggtg
     1321 tgactatcca aattatgact gagcaggagt caggatactg ctggagaaac tttgtgaatt
     1381 atagcccgag taatgaagcc cactggccta ggtatcccca tctgtgggta cgactgtacg
     1441 ttcttgaact gtactgcatc atactgggcc tgcctccttg tctcaacatt ctgagaagga
     1501 agcagccaca gctgacattc tttaccatcg ctcttcagtc ttgtcattac cagcgactgc
     1561 ccccacacat tctctgggcc accgggttga aaagcggcag cgagactccc gggacctcag
     1621 agtccgccac acccgaaagt gataaaaagt attctattgg tttagccatc ggcactaatt
     1681 ccgttggatg ggctgtcata accgatgaat acaaagtacc ttcaaagaaa tttaaggtgt
     1741 tggggaacac agaccgtcat tcgattaaaa agaatcttat cggtgccctc ctattcgata
     1801 gtggcgaaac ggcagaggcg actcgcctga aacgaaccgc tcggagaagg tatacacgtc
     1861 gcaagaaccg aatatgttac ttacaagaaa tttttagcaa tgagatggcc aaagttgacg
     1921 attctttctt tcaccgtttg gaagagtcct tccttgtcga agaggacaag aaacatgaac
     1981 ggcaccccat ctttggaaac atagtagatg aggtggcata tcatgaaaag tacccaacga
     2041 tttatcacct cagaaaaaag ctagttgact caactgataa agcggacctg aggttaatct
     2101 acttggctct tgcccatatg ataaagttcc gtgggcactt tctcattgag ggtgatctaa
     2161 atccggacaa ctcggatgtc gacaaactgt tcatccagtt agtacaaacc tataatcagt
     2221 tgtttgaaga gaaccctata aatgcaagtg gcgtggatgc gaaggctatt cttagcgccc
     2281 gcctctctaa atcccgacgg ctagaaaacc tgatcgcaca attacccgga gagaagaaaa
     2341 atgggttgtt cggtaacctt atagcgctct cactaggcct gacaccaaat tttaagtcga
     2401 acttcgactt agctgaagat gccaaattgc agcttagtaa ggacacgtac gatgacgatc
     2461 tcgacaatct actggcacaa attggagatc agtatgcgga cttatttttg gctgccaaaa
     2521 accttagcga tgcaatcctc ctatctgaca tactgagagt taatactgag attaccaagg
     2581 cgccgttatc cgcttcaatg atcaaaaggt acgatgaaca tcaccaagac ttgacacttc
     2641 tcaaggccct agtccgtcag caactgcctg agaaatataa ggaaatattc tttgatcagt
     2701 cgaaaaacgg gtacgcaggt tatattgacg gcggagcgag tcaagaggaa ttctacaagt
     2761 ttatcaaacc catattagag aagatggatg ggacggaaga gttgcttgta aaactcaatc
     2821 gcgaagatct actgcgaaag cagcggactt tcgacaacgg tagcattcca catcaaatcc
     2881 acttaggcga attgcatgct atacttagaa ggcaggagga tttttatccg ttcctcaaag
     2941 acaatcgtga aaagattgag aaaatcctaa cctttcgcat accttactat gtgggacccc
     3001 tggcccgagg gaactctcgg ttcgcatgga tgacaagaaa gtccgaagaa acgattactc
     3061 catggaattt tgaggaagtt gtcgataaag gtgcgtcagc tcaatcgttc atcgagagga
     3121 tgaccaactt tgacaagaat ttaccgaacg aaaaagtatt gcctaagcac agtttacttt
     3181 acgagtattt cacagtgtac aatgaactca cgaaagttaa gtatgtcact gagggcatgc
     3241 gtaaacccgc ctttctaagc ggagaacaga agaaagcaat agtagatctg ttattcaaga
     3301 ccaaccgcaa agtgacagtt aagcaattga aagaggacta ctttaagaaa attgaatgct
     3361 tcgattctgt cgagatctcc ggggtagaag atcgatttaa tgcgtcactt ggtacgtatc
     3421 atgacctcct aaagataatt aaagataagg acttcctgga taacgaagag aatgaagata
     3481 tcttagaaga tatagtgttg actcttaccc tctttgaaga tcgggaaatg attgaggaaa
     3541 gactaaaaac atacgctcac ctgttcgacg ataaggttat gaaacagtta aagaggcgtc
     3601 gctatacggg ctggggacga ttgtcgcgga aacttatcaa cgggataaga gacaagcaaa
     3661 gtggtaaaac tattctcgat tttctaaaga gcgacggctt cgccaatagg aactttatgc
     3721 agctgatcca tgatgactct ttaaccttca aagaggatat acaaaaggca caggtttccg
     3781 gacaagggga ctcattgcac gaacatattg cgaatcttgc tggttcgcca gccatcaaaa
     3841 agggcatact ccagacagtc aaagtagtgg atgagctagt taaggtcatg ggacgtcaca
     3901 aaccggaaaa cattgtaatc gagatggcac gcgaaaatca aacgactcag aaggggcaaa
     3961 aaaacagtcg agagcggatg aagagaatag aagagggtat taaagaactg ggcagccaga
     4021 tcttaaagga gcatcctgtg gaaaataccc aattgcagaa cgagaaactt tacctctatt
     4081 acctacaaaa tggaagggac atgtatgttg atcaggaact ggacataaac cgtttatctg
     4141 attacgacgt cgatcacatt gtaccccaat cctttttgaa ggacgattca atcgacaata
     4201 aagtgcttac acgctcggat aagaaccgag ggaaaagtga caatgttcca agcgaggaag
     4261 tcgtaaagaa aatgaagaac tattggcggc agctcctaaa tgcgaaactg ataacgcaaa
     4321 gaaagttcga taacttaact aaagctgaga ggggtggctt gtctgaactt gacaaggccg
     4381 gatttattaa acgtcagctc gtggaaaccc gccaaatcac aaagcatgtt gcacagatac
     4441 tagattcccg aatgaatacg aaatacgacg agaacgataa gctgattcgg gaagtcaaag
     4501 taatcacttt aaagtcaaaa ttggtgtcgg acttcagaaa ggattttcaa ttctataaag
     4561 ttagggagat aaataactac caccatgcgc acgacgctta tcttaatgcc gtcgtaggga
     4621 ccgcactcat taagaaatac ccgaagctag aaagtgagtt tgtgtatggt gattacaaag
     4681 tttatgacgt ccgtaagatg atcgcgaaaa gcgaacagga gataggcaag gctacagcca
     4741 aatacttctt ttattctaac attatgaatt tctttaagac ggaaatcact ctggcaaacg
     4801 gagagatacg caaacgacct ttaattgaaa ccaatgggga gacaggtgaa atcgtatggg
     4861 ataagggccg ggacttcgcg acggtgagaa aagttttgtc catgccccaa gtcaacatag
     4921 taaagaaaac tgaggtgcag accggagggt tttcaaagga atcgattctt ccaaaaagga
     4981 atagtgataa gctcatcgct cgtaaaaagg actgggaccc gaaaaagtac ggtggcttcg
     5041 atagccctac agttgcctat tctgtcctag tagtggcaaa agttgagaag ggaaaatcca
     5101 agaaactgaa gtcagtcaaa gaattattgg ggataacgat tatggagcgc tcgtcttttg
     5161 aaaagaaccc catcgacttc cttgaggcga aaggttacaa ggaagtaaaa aaggatctca
     5221 taattaaact accaaagtat agtctgtttg agttagaaaa tggccgaaaa cggatgttgg
     5281 ctagcgccgg agagcttcaa aaggggaacg aactcgcact accgtctaaa tacgtgaatt
     5341 tcctgtattt agcgtcccat tacgagaagt tgaaaggttc acctgaagat aacgaacaga
     5401 agcaactttt tgttgagcag cacaaacatt atctcgacga aatcatagag caaatttcgg
     5461 aattcagtaa gagagtcatc ctagctgatg ccaatctgga caaagtatta agcgcataca
     5521 acaagcacag ggataaaccc atacgtgagc aggcggaaaa tattatccat ttgtttactc
     5581 ttaccaacct cggcgctcca gccgcattca agtattttga cacaacgata gatcgcaaac
     5641 gatacacttc taccaaggag gtgctagacg cgacactgat tcaccaatcc atcacgggat
     5701 tatatgaaac tcggatagat ttgtcacagc ttgggggtga ctctggtggt tctactaatc
     5761 tgtcagatat tattgaaaag gagaccggta agcaactggt tatccaggaa tccatcctca
     5821 tgctcccaga ggaggtggaa gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc
     5881 acaccgccta cgacgagagc accgacgaga atgtcatgct tctgactagc gacgcccctg
     5941 aatacaagcc ttgggctctg gtcatacagg atagcaacgg tgagaacaag attaagatgc
     6001 tctctggtgg ttctcccaag aagaagagga aagtcgaaga tcctaagaaa aagcggaaag
     6061 tgtaacgcgt gctagcttcg aagctaattc actcctcagg tgcaggctgc ctatcagaag
     6121 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     6181 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     6241 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     6301 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     6361 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     6421 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     6481 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     6541 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     6601 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     6661 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6721 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6781 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc gatctgacaa tgttcagtgc
     6841 agagactcgg ctacgcctcg tggactttga agttgaccaa caatgtttat tcttacctct
     6901 aatagtcctc tgtggcaagg tcaagattct gttagaagcc aatgaagaac ctggttgttc
     6961 aataacattt tgttcgtcta atatttcact accgcttgac gttggctgca cttcatgtac
     7021 ctcatctata aacgcttctt ctgtatcgct ctggacgtca tcttcactta cgtgatctga
     7081 tatttcactg tcagaatcct caccaacaag ctcgtcatcg ctttgcagaa gagcagagag
     7141 gatatgctca tcgtctaaag aactacccat tttattatat attagtcacg atatctataa
     7201 caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt
     7261 atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac
     7321 gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga
     7381 gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa
     7441 gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt
     7501 aaaaaagatt tgcgctttac tcgacctaaa ctttaaacac gtcatagaat cttcgtttga
     7561 caaaaaccac attgtggcca agctgtgtga cgcgacgcgc gctaaagaat ggcaaaccaa
     7621 gtcgcgcgag cgtcgacctc gagggggggc ccggtaccca gcttttgttc cctttagtga
     7681 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     8581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9841 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     9901 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     9961 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
    10021 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
    10081 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
    10141 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
    10201 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
    10261 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
    10321 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
    10381 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
    10441 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
    10501 agggcgaatt ggagctccac cgcggccgcc cggtttatcg ttaatatgga tcaatttgaa
    10561 cagttgatta acgtgtctct gctcaagtct ttgatcaaaa cgcaaatcga cgaaaatgtg
    10621 tcggacaata tcaagtcgat gagcgaaaaa ctaaaaaggc tagaatacga caatctcaca
    10681 gacagcgttg agatatacgg tattcacgac agcaggctga ataataaaaa aattagaaac
    10741 tattatttaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgcgta
    10801 aaattgacgc atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga
    10861 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt
    10921 tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa
    10981 aacttttaaa cattctctct tttacaaaaa taaacttatt ttgtacttta aaaacagtca
    11041 tgttgtatta taaaataagt aattagctta acttatacat aatagaaaca aattatactt
    11101 attagtcagt cagaaacaac tttggcacat atcaatatta tgctctcgac aaataacttt
    11161 tttgcatttt ttgcacgatg catttgcctt tcgccttatt ttagaggggc agtaagtaca
    11221 gtaagtacgt tttttcatta ctggctcttc agtactgtca tctgatgtac caggcacttc
    11281 atttggcaaa atattagaga tattatcgcg caaatatctc ttcaaagtag gagcttctaa
    11341 acgcttacgc ataaacgatg acgtcaggct catgtaaagg tttctcataa attttttgcg
    11401 actttgaacc ttttctccct tgctactgac attatggctg tatataataa aagaatttat
    11461 gcaggcaatg tttatcattc cgtacaataa tgccataggc cacctattcg tcttcctact
    11521 gcaggtcatc acagaacaca tttggtctag cgtgtccact ccgcctttag tttgattata
    11581 atacataacc atttgcggtt taccggtact ttcgttgata gaagcatcct catcacaaga
    11641 tgataataag tataccatct tagctggctt cggtttatat gagacgagag taaggggtcc
    11701 gtcaaaacaa aacatcgatg ttcccactgg cctggagcga ctgtttttca gtacttccgg
    11761 tatctcgcgt ttgtttgatc gcacggttcc cacaatggtt aattcgagct cgcccaaacc
    11821 gggcgcgccg ggcatttaaa tggaattcga tgccctatat atggatccaa ttgcaatgat
    11881 catcatgaca gatctcgaca ttgattattg actagttatt aatagtaatc aattacgggg
    11941 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
    12001 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
    12061 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
    12121 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
    12181 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
    12241 cagtacatct acgtattagt catcgctatt accatg
//
