LOCUS       sequence-413618-       13049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing doxycycline-inducible human 
            codon-optimized adenine base editor where TadA8.20m is fused to the 
            N-terminus of SpCas9(D10A)-6xNLS .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13049)
  AUTHORS   Xiao MS, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, 
            King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson 
            T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
  TITLE     Genome-scale exon perturbation screens uncover exons critical for 
            cell fitness.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2553-2572.e19. doi: 
            10.1016/j.molcel.2024.05.024. Epub 2024 Jun 24.
  PUBMED    38917794
REFERENCE   2  (bases 1 to 13049)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2024.05.024"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2553-2572.e19"
COMMENT     Sequence Label: pLenti-nSpCas9-TadA8.20m
FEATURES             Location/Qualifiers
     source          1..13049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..86
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    287..404
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        469..783
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    477..495
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    513..531
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    548..566
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    584..602
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    620..638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    655..673
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    691..709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     756..780
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             806..829
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             830..856
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             863..1360
                     /codon_start=1
                     /product="DNA adenosine deaminase evolved from the E. coli 
                     tRNA adenosine deaminase TadA (Gaudelli et al., 2017)"
                     /label=TadA*(7.10)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLYDATLYSTFEPCVMCAGAMIHSRIGRVVFG
                     VRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLCRFFRMPRRVFNAQKKAQS
                     STD"
     CDS             join(1463..1486,1487..1489,1490..5563)
                     /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             1463..5563
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="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             5585..5611
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5621..5647
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5657..5683
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5693..5719
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5729..5755
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5765..5791
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     regulatory      5801..5810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        5810..6021
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6037..6432
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             6439..6492
                     /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             join(6493..7110,7111..7116,7117..7152,7153..7155,7156..7191,
                     7192..7194,7195..7230,7231..7239)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    7255..7843
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7308..7328)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7726..7737)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7915..8148
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8226..8347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8263..8282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8317..8336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8387..8522
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8542..8561)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8543..8561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8571..8588)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8571..8587)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8580..8602)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8729..9184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8816..8835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9026..9047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9210..9314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9315..9383,9384..10175)
                     /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     complement(9533..9552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10346..10934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10835..10854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11088..11105
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11222..11243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11258..11263,11264..11281,11282..11288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11296..11312
                     /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)."
     primer_bind     11301..11323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11320..11336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11320..11336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11355..11375
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11357..11375
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11403..11629
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11630..11810
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11857..11982
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12475..12708
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12893..12937
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
       61 ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
      121 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      181 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      241 atcataaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      301 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      361 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      421 acagcagaga tccactttgg cgccggctcg agcgcgaatc gatatgtcga gtttactccc
      481 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg
      541 agtttactcc ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga
      601 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta
      661 tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
      721 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
      781 gcctggagaa ttggctagca ccatggatta caaagacgat gacgataagt acccatacga
      841 tgtgccagat tacgccagcg cttccgaagt cgagttttcc catgagtact ggatgagaca
      901 cgcattgact ctcgcaaaga gggctcgaga tgaacgcgag gtgcccgtgg gggcagtact
      961 cgtgctcaac aatcgcgtaa tcggcgaagg ttggaatagg gcaatcggac tccacgaccc
     1021 cactgcacat gcggaaatca tggcccttcg acagggaggg cttgtgatgc agaattatcg
     1081 actttatgat gcgacgctgt actcgacgtt tgaaccttgc gtaatgtgcg cgggagctat
     1141 gattcactcc cgcattggac gagttgtatt cggtgttcgc aacgccaaga cgggtgccgc
     1201 aggttcactg atggacgtgc tgcatcatcc aggcatgaac caccgggtag aaatcacaga
     1261 aggcatattg gcggacgaat gtgcggcgct gttgtgtcgt ttttttcgca tgcccaggcg
     1321 ggtctttaac gcccagaaaa aagcacaatc ctctactgac tctggtggtt cttctggtgg
     1381 ttctagcggc agcgagactc ccgggacctc agagtccgcc acacccgaaa gttctggtgg
     1441 ttcttctggt ggttctagcg ctgacaagaa gtacagcatc ggcctggcca tcggcaccaa
     1501 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     1561 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     1621 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     1681 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     1741 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     1801 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     1861 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     1921 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     1981 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     2041 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     2101 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     2161 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     2221 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     2281 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     2341 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     2401 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     2461 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     2521 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     2581 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     2641 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     2701 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     2761 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     2821 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     2881 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     2941 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     3001 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     3061 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     3121 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     3181 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     3241 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     3301 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     3361 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     3421 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     3481 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     3541 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     3601 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     3661 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     3721 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     3781 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     3841 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     3901 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     3961 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     4021 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     4081 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     4141 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     4201 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     4261 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     4321 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     4381 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     4441 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     4501 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     4561 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     4621 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     4681 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     4741 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     4801 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     4861 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     4921 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     4981 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     5041 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     5101 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     5161 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     5221 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     5281 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     5341 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     5401 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     5461 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     5521 cctgtacgag acacggatcg acctgtctca gctgggaggt gacagcggcg ggaattccag
     5581 atcccctgct gctaaacgtg ttaagcttga tgggggtagc ccggcagcca agagagtcaa
     5641 actcgacggc ggctccccag cggcaaaaag ggtgaaacta gacgggggta gccccgccgc
     5701 gaagcgtgta aagctggatg gaggatcgcc tgcggctaag cgagtcaaat tagacggtgg
     5761 gtcaccggct gcaaagcgcg tcaaattgga ttgagctagc gccaccatgg ggcagagcgc
     5821 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag
     5881 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     5941 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     6001 gggtttgccg ccagaacaca gttcgaacaa ttcaccatgg ccaagccttt gtctcaagaa
     6061 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     6121 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     6181 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     6241 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     6301 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     6361 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     6421 gtgtgggagg gcggtaccga aggtagaggt tctctcctca cttgtggtga tgttgaagaa
     6481 aaccctggtc caatgtctag actggacaag agcaaagtca taaacggagc tctggaatta
     6541 ctcaatggtg tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt
     6601 gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca
     6661 atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa
     6721 gactttctgc ggaacaacgc caagtcatac cgctgtgctc tcctctcaca tcgcgacggg
     6781 gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag
     6841 ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc
     6901 gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag
     6961 gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg
     7021 ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc
     7081 ctggagaaac agctaaagtg cgaaagcggc gggccgaccg acgcccttga cgattttgac
     7141 ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac
     7201 gctcttgacg attttgacct tgacatgctc cccgggtaaa cgcgttaagt cgacaatcaa
     7261 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     7321 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     7381 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     7441 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     7501 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     7561 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     7621 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     7681 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     7741 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     7801 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     7861 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     7921 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     7981 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     8041 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     8101 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     8161 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     8221 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     8281 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     8341 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     8401 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     8461 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     8521 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     8581 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     8641 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     8701 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     8761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     9001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     9061 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     9121 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     9181 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     9241 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     9301 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     9361 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     9421 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     9481 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     9541 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     9601 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     9661 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     9721 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     9781 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     9841 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     9901 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     9961 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
    10021 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
    10081 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
    10141 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
    10201 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
    10261 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
    10321 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
    10381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
    10441 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
    10501 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
    10561 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
    10621 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
    10681 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
    10741 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
    10801 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
    10861 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
    10921 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
    10981 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
    11041 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
    11101 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
    11161 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
    11221 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
    11281 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
    11341 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
    11401 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
    11461 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
    11521 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
    11581 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
    11641 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    11701 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    11761 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
    11821 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
    11881 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
    11941 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
    12001 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
    12061 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
    12121 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
    12181 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
    12241 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
    12301 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
    12361 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
    12421 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
    12481 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
    12541 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
    12601 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
    12661 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
    12721 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
    12781 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
    12841 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
    12901 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
    12961 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
    13021 agtttttgct gtactttcta tagtgaata
//
