LOCUS       sequence-208397-       12505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for dox-inducible expression of BE3 (not codon 
            optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12505)
  AUTHORS   Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
            Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, 
            Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow 
            LE
  TITLE     Optimized base editors enable efficient editing in cells, organoids 
            and mice.
  JOURNAL   Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
  PUBMED    29969439
REFERENCE   2  (bases 1 to 12505)
  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 
            Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194."
COMMENT     Sequence Label: pLenti-TRE3G-BE3-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2623..3001
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2631..2649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2667..2685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2703..2721
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2739..2757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2775..2793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2811..2829
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2847..2865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2910..2934
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3028..3714
                     /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(3763..7161,7162..7164,7165..7761,7762,7763..7765,
                     7766..7767,7768..7770,7771..7863)
                     /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(3763..3786,3787..3789,3790..7863)
                     /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             7876..8124
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8137..8157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8166..8183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        8222..8721
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8257..8280)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8654..8673
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8742..9341
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8742..8761)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9238..9258
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9394..9982
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9447..9467)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9865..9876)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9985..10001)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9986..10002)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10511..10691
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(10753..11341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10833..10852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11512..12303,12304..12372))
                     /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     12135..12154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12373..12477)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc gattctagat tcgagtttac tccctatcag
     2641 tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta tgcagacttt
     2701 actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg atagagaacg
     2761 tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac tccctatcag
     2821 tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta taagctttag
     2881 gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg
     2941 agcaattcca caacactttt gtcttatacc aactttccgt accacttcct accctcgtaa
     3001 agtcgagctt gcgttggatc cgccaccatg agctcagaga ctggcccagt ggctgtggac
     3061 cccacattga gacggcggat cgagccccat gagtttgagg tattcttcga tccgagagag
     3121 ctccgcaagg agacctgcct gctttacgaa attaattggg ggggccggca ctccatttgg
     3181 cgacatacat cacagaacac taacaagcac gtcgaagtca acttcatcga gaagttcacg
     3241 acagaaagat atttctgtcc gaacacaagg tgcagcatta cctggtttct cagctggagc
     3301 ccatgcggcg aatgtagtag ggccatcact gaattcctgt caaggtatcc ccacgtcact
     3361 ctgtttattt acatcgcaag gctgtaccac cacgctgacc cccgcaatcg acaaggcctg
     3421 cgggatttga tctcttcagg tgtgactatc caaattatga ctgagcagga gtcaggatac
     3481 tgctggagaa actttgtgaa ttatagcccg agtaatgaag cccactggcc taggtatccc
     3541 catctgtggg tacgactgta cgttcttgaa ctgtactgca tcatactggg cctgcctcct
     3601 tgtctcaaca ttctgagaag gaagcagcca cagctgacat tctttaccat cgctcttcag
     3661 tcttgtcatt accagcgact gcccccacac attctctggg ccaccgggtt gaaaagcggc
     3721 agcgagactc ccgggacctc agagtccgcc acacccgaaa gtgataaaaa gtattctatt
     3781 ggtttagcca tcggcactaa ttccgttgga tgggctgtca taaccgatga atacaaagta
     3841 ccttcaaaga aatttaaggt gttggggaac acagaccgtc attcgattaa aaagaatctt
     3901 atcggtgccc tcctattcga tagtggcgaa acggcagagg cgactcgcct gaaacgaacc
     3961 gctcggagaa ggtatacacg tcgcaagaac cgaatatgtt acttacaaga aatttttagc
     4021 aatgagatgg ccaaagttga cgattctttc tttcaccgtt tggaagagtc cttccttgtc
     4081 gaagaggaca agaaacatga acggcacccc atctttggaa acatagtaga tgaggtggca
     4141 tatcatgaaa agtacccaac gatttatcac ctcagaaaaa agctagttga ctcaactgat
     4201 aaagcggacc tgaggttaat ctacttggct cttgcccata tgataaagtt ccgtgggcac
     4261 tttctcattg agggtgatct aaatccggac aactcggatg tcgacaaact gttcatccag
     4321 ttagtacaaa cctataatca gttgtttgaa gagaacccta taaatgcaag tggcgtggat
     4381 gcgaaggcta ttcttagcgc ccgcctctct aaatcccgac ggctagaaaa cctgatcgca
     4441 caattacccg gagagaagaa aaatgggttg ttcggtaacc ttatagcgct ctcactaggc
     4501 ctgacaccaa attttaagtc gaacttcgac ttagctgaag atgccaaatt gcagcttagt
     4561 aaggacacgt acgatgacga tctcgacaat ctactggcac aaattggaga tcagtatgcg
     4621 gacttatttt tggctgccaa aaaccttagc gatgcaatcc tcctatctga catactgaga
     4681 gttaatactg agattaccaa ggcgccgtta tccgcttcaa tgatcaaaag gtacgatgaa
     4741 catcaccaag acttgacact tctcaaggcc ctagtccgtc agcaactgcc tgagaaatat
     4801 aaggaaatat tctttgatca gtcgaaaaac gggtacgcag gttatattga cggcggagcg
     4861 agtcaagagg aattctacaa gtttatcaaa cccatattag agaagatgga tgggacggaa
     4921 gagttgcttg taaaactcaa tcgcgaagat ctactgcgaa agcagcggac tttcgacaac
     4981 ggtagcattc cacatcaaat ccacttaggc gaattgcatg ctatacttag aaggcaggag
     5041 gatttttatc cgttcctcaa agacaatcgt gaaaagattg agaaaatcct aacctttcgc
     5101 ataccttact atgtgggacc cctggcccga gggaactctc ggttcgcatg gatgacaaga
     5161 aagtccgaag aaacgattac tccatggaat tttgaggaag ttgtcgataa aggtgcgtca
     5221 gctcaatcgt tcatcgagag gatgaccaac tttgacaaga atttaccgaa cgaaaaagta
     5281 ttgcctaagc acagtttact ttacgagtat ttcacagtgt acaatgaact cacgaaagtt
     5341 aagtatgtca ctgagggcat gcgtaaaccc gcctttctaa gcggagaaca gaagaaagca
     5401 atagtagatc tgttattcaa gaccaaccgc aaagtgacag ttaagcaatt gaaagaggac
     5461 tactttaaga aaattgaatg cttcgattct gtcgagatct ccggggtaga agatcgattt
     5521 aatgcgtcac ttggtacgta tcatgacctc ctaaagataa ttaaagataa ggacttcctg
     5581 gataacgaag agaatgaaga tatcttagaa gatatagtgt tgactcttac cctctttgaa
     5641 gatcgggaaa tgattgagga aagactaaaa acatacgctc acctgttcga cgataaggtt
     5701 atgaaacagt taaagaggcg tcgctatacg ggctggggac gattgtcgcg gaaacttatc
     5761 aacgggataa gagacaagca aagtggtaaa actattctcg attttctaaa gagcgacggc
     5821 ttcgccaata ggaactttat gcagctgatc catgatgact ctttaacctt caaagaggat
     5881 atacaaaagg cacaggtttc cggacaaggg gactcattgc acgaacatat tgcgaatctt
     5941 gctggttcgc cagccatcaa aaagggcata ctccagacag tcaaagtagt ggatgagcta
     6001 gttaaggtca tgggacgtca caaaccggaa aacattgtaa tcgagatggc acgcgaaaat
     6061 caaacgactc agaaggggca aaaaaacagt cgagagcgga tgaagagaat agaagagggt
     6121 attaaagaac tgggcagcca gatcttaaag gagcatcctg tggaaaatac ccaattgcag
     6181 aacgagaaac tttacctcta ttacctacaa aatggaaggg acatgtatgt tgatcaggaa
     6241 ctggacataa accgtttatc tgattacgac gtcgatcaca ttgtacccca atcctttttg
     6301 aaggacgatt caatcgacaa taaagtgctt acacgctcgg ataagaaccg agggaaaagt
     6361 gacaatgttc caagcgagga agtcgtaaag aaaatgaaga actattggcg gcagctccta
     6421 aatgcgaaac tgataacgca aagaaagttc gataacttaa ctaaagctga gaggggtggc
     6481 ttgtctgaac ttgacaaggc cggatttatt aaacgtcagc tcgtggaaac ccgccaaatc
     6541 acaaagcatg ttgcacagat actagattcc cgaatgaata cgaaatacga cgagaacgat
     6601 aagctgattc gggaagtcaa agtaatcact ttaaagtcaa aattggtgtc ggacttcaga
     6661 aaggattttc aattctataa agttagggag ataaataact accaccatgc gcacgacgct
     6721 tatcttaatg ccgtcgtagg gaccgcactc attaagaaat acccgaagct agaaagtgag
     6781 tttgtgtatg gtgattacaa agtttatgac gtccgtaaga tgatcgcgaa aagcgaacag
     6841 gagataggca aggctacagc caaatacttc ttttattcta acattatgaa tttctttaag
     6901 acggaaatca ctctggcaaa cggagagata cgcaaacgac ctttaattga aaccaatggg
     6961 gagacaggtg aaatcgtatg ggataagggc cgggacttcg cgacggtgag aaaagttttg
     7021 tccatgcccc aagtcaacat agtaaagaaa actgaggtgc agaccggagg gttttcaaag
     7081 gaatcgattc ttccaaaaag gaatagtgat aagctcatcg ctcgtaaaaa ggactgggac
     7141 ccgaaaaagt acggtggctt cgatagccct acagttgcct attctgtcct agtagtggca
     7201 aaagttgaga agggaaaatc caagaaactg aagtcagtca aagaattatt ggggataacg
     7261 attatggagc gctcgtcttt tgaaaagaac cccatcgact tccttgaggc gaaaggttac
     7321 aaggaagtaa aaaaggatct cataattaaa ctaccaaagt atagtctgtt tgagttagaa
     7381 aatggccgaa aacggatgtt ggctagcgcc ggagagcttc aaaaggggaa cgaactcgca
     7441 ctaccgtcta aatacgtgaa tttcctgtat ttagcgtccc attacgagaa gttgaaaggt
     7501 tcacctgaag ataacgaaca gaagcaactt tttgttgagc agcacaaaca ttatctcgac
     7561 gaaatcatag agcaaatttc ggaattcagt aagagagtca tcctagctga tgccaatctg
     7621 gacaaagtat taagcgcata caacaagcac agggataaac ccatacgtga gcaggcggaa
     7681 aatattatcc atttgtttac tcttaccaac ctcggcgctc cagccgcatt caagtatttt
     7741 gacacaacga tagatcgcaa acgatacact tctaccaagg aggtgctaga cgcgacactg
     7801 attcaccaat ccatcacggg attatatgaa actcggatag atttgtcaca gcttgggggt
     7861 gactctggtg gttctactaa tctgtcagat attattgaaa aggagaccgg taagcaactg
     7921 gttatccagg aatccatcct catgctccca gaggaggtgg aagaagtcat tgggaacaag
     7981 ccggaaagcg atatactcgt gcacaccgcc tacgacgaga gcaccgacga gaatgtcatg
     8041 cttctgacta gcgacgcccc tgaatacaag ccttgggctc tggtcataca ggatagcaac
     8101 ggtgagaaca agattaagat gctctctggt ggttctccca agaagaagag gaaagtctaa
     8161 ccggtcatca tcaccatcac cattgaaaac cgctgttcct aggaatcccg aggcctctac
     8221 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
     8281 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
     8341 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
     8401 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
     8461 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
     8521 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
     8581 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
     8641 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
     8701 tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca
     8761 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     8821 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     8881 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     8941 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     9001 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     9061 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     9121 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     9181 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     9241 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     9301 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaaacgttgt tgtttggggt
     9361 tgaattactc ttctagaccg cggcgcgccc gataatcaac ctctggatta caaaatttgt
     9421 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     9481 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     9541 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     9601 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     9661 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     9721 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     9781 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     9841 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     9901 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     9961 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
    10021 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
    10081 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
    10141 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    10201 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
    10261 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
    10321 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
    10381 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
    10441 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
    10501 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    10561 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    10621 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    10681 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    10741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    10801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    10861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    10921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    10981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
    11041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
    11101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    11161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    11221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    11281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    11341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    11401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    11461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    11521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    11581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    11641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    11701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    11761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    11821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    11881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    11941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    12001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    12061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    12121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    12181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    12241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    12301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    12361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    12421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    12481 atttccccga aaagtgccac ctgac
//
