LOCUS       sequence-413615-       14156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing doxycycline-inducible human 
            codon-optimized cytosine base editor where evoCDA1 is fused to the 
            N-terminus of SpCas9(D10A)-sDNA Rad51-2xUGI-6xNLS .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14156)
  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 14156)
  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-evoCDA1
FEATURES             Location/Qualifiers
     source          1..14156
                     /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             join(2024..2047,2048..2050,2051..6124)
                     /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             2024..6124
                     /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             6161..6409
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6440..6688
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6713..6739
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6749..6775
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6785..6811
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6821..6847
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6857..6883
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6893..6919
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     regulatory      6929..6938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        6938..7149
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7165..7560
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             7567..7620
                     /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(7621..8238,8239..8244,8245..8280,8281..8283,8284..8319,
                     8320..8322,8323..8358,8359..8367)
                     /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    8383..8971
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8436..8456)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8854..8865)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9043..9276
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9354..9475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9391..9410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9445..9464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9494..9629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9649..9668)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9650..9668)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9678..9695)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9678..9694)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9687..9709)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9836..10291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9923..9942)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10133..10154
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10317..10421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10422..10490,10491..11282)
                     /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(10640..10659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11453..12041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11942..11961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12195..12212
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12329..12350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12365..12370,12371..12388,12389..12395)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12403..12419
                     /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     12408..12430
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12427..12443
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12427..12443
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12462..12482
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12464..12482
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        12510..12736
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             12737..12917
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12964..13089
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    13582..13815
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             14000..14044
                     /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 ctaccgacgc cgagtacgtg cggatccacg agaagctgga
      901 tatctataca ttcaagaagc agtttagcaa caataagaag tccgtgtctc acagatgcta
      961 cgtgctgttc gagctgaagc ggagaggaga gaggcgcgcc tgtttttggg gctatgccgt
     1021 gaacaagcca cagtctggaa ccgagagggg aatccacgca gagatcttca gcatcaggaa
     1081 ggtggaggag tacctgcgcg acaaccccgg ccagtttaca atcaattggt atagctcctg
     1141 gagcccttgc gccgattgtg ccgagaagat cctggagtgg tacaaccagg agctgagggg
     1201 caatggccac accctgaaga tctgggtgtg caagctgtac tatgagaaga acgccaggaa
     1261 tcagatcggc ctgtggaacc tgcgcgacaa tggcgtgggc ctgaacgtga tggtgtccga
     1321 gcactatcag tgctgtcgca agatctttat ccagtctagc cacaatcagc tgaacgagaa
     1381 tcggtggctg gagaagacac tgaagagagc cgagaagcgg agaagcgagc tgtccatcat
     1441 gtttcaggtg aagatcctgc acaccacaaa gtctcccgcc gtgagcggag gatctagcgg
     1501 aggatcaagc ggaagcgaga ctcctggaac cagcgaaagc gcaaccccag aaagcagcgg
     1561 aggaagtagc ggaggaagca tggcaatgca gatgcagctt gaagcaaatg cagatacttc
     1621 agtggaagaa gaaagctttg gcccacaacc catttcacgg ttagagcagt gtggcataaa
     1681 tgccaacgat gtgaagaaat tggaagaagc tggattccat actgtggagg ctgttgccta
     1741 tgcgccaaag aaggagctaa taaatattaa gggaattagt gaagccaaag ctgataaaat
     1801 tctggctgag gcagctaaat tagttccaat gggtttcacc actgcaactg aattccacca
     1861 aaggcggtca gagatcatac agattactac tggctccaaa gagcttgaca aactacttca
     1921 atctggcgga tctagcggtg gatctagcgg ctctgagacc cctggaacat ccgaatccgc
     1981 cactccagag agcagcggag gctcttctgg aggatcaagc gctgacaaga agtacagcat
     2041 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     2101 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     2161 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     2221 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     2281 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     2341 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     2401 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     2461 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     2521 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     2581 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     2641 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     2701 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
     2761 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     2821 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     2881 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     2941 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     3001 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     3061 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     3121 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     3181 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     3241 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     3301 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     3361 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     3421 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     3481 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     3541 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     3601 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     3661 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     3721 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     3781 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     3841 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     3901 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     3961 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     4021 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     4081 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     4141 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     4201 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     4261 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     4321 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     4381 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     4441 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     4501 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
     4561 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
     4621 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     4681 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     4741 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     4801 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     4861 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     4921 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     4981 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     5041 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     5101 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     5161 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     5221 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     5281 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     5341 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     5401 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     5461 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     5521 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     5581 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     5641 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     5701 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     5761 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     5821 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     5881 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     5941 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     6001 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     6061 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     6121 tgacagtggc ggaagtggac tagtaggaag tggcgggagc actaatctga gcgacatcat
     6181 tgagaaggag actgggaaac agctggtcat tcaggagtcc atcctgatgc tgcctgagga
     6241 ggtggaggaa gtgatcggca acaagccaga gtctgacatc ctggtgcaca ccgcctacga
     6301 cgagtccaca gatgagaatg tgatgctgct gacctctgac gcccccgagt ataagccttg
     6361 ggccctggtc atccaggatt ctaacggcga gaataagatc aagatgctga gcggaggatc
     6421 cggaggatct ggaggcagca ccaacctgtc tgacatcatc gagaaggaga caggcaagca
     6481 gctggtcatc caggagagca tcctgatgct gcccgaagaa gtcgaagaag tgatcggaaa
     6541 caagcctgag agcgatatcc tggtccatac cgcctacgac gagagtaccg acgaaaatgt
     6601 gatgctgctg acatccgacg ccccagagta taagccctgg gctctggtca tccaggattc
     6661 caacggagag aacaaaatca aaatgctgtc tggcggctca gaattcggat cccctgctgc
     6721 taaacgtgtt aagcttgatg ggggtagccc ggcagccaag agagtcaaac tcgacggcgg
     6781 ctccccagcg gcaaaaaggg tgaaactaga cgggggtagc cccgccgcga agcgtgtaaa
     6841 gctggatgga ggatcgcctg cggctaagcg agtcaaatta gacggtgggt caccggctgc
     6901 aaagcgcgtc aaattggatt gagctagcgc caccatgggg cagagcgcac atcgcccaca
     6961 gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc
     7021 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     7081 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     7141 agaacacagt tcgaacaatt caccatggcc aagcctttgt ctcaagaaga atccaccctc
     7201 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     7261 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     7321 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     7381 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     7441 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     7501 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     7561 ggtaccgaag gtagaggttc tctcctcact tgtggtgatg ttgaagaaaa ccctggtcca
     7621 atgtctagac tggacaagag caaagtcata aacggagctc tggaattact caatggtgtc
     7681 ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga gcagcctacc
     7741 ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat cgagatgctg
     7801 gacaggcatc atacccactt ctgccccctg gaaggcgagt catggcaaga ctttctgcgg
     7861 aacaacgcca agtcataccg ctgtgctctc ctctcacatc gcgacggggc taaagtgcat
     7921 ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg
     7981 tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt gggccacttt
     8041 acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga aagagagaca
     8101 cctaccaccg attctatgcc cccacttctg agacaagcaa ttgagctgtt cgaccggcag
     8161 ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct ggagaaacag
     8221 ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg attttgactt agacatgctc
     8281 ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc tcttgacgat
     8341 tttgaccttg acatgctccc cgggtaaacg cgttaagtcg acaatcaacc tctggattac
     8401 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     8461 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     8521 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     8581 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     8641 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     8701 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     8761 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     8821 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     8881 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     8941 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     9001 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     9061 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     9121 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     9181 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     9241 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     9301 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     9361 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     9421 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     9481 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     9541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     9601 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     9661 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     9721 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     9781 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     9841 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     9901 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9961 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    10021 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    10081 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    10141 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
    10201 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    10261 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
    10321 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    10381 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    10441 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    10501 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    10561 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    10621 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
    10681 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    10741 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    10801 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    10861 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    10921 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    10981 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    11041 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    11101 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    11161 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    11221 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    11281 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    11341 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    11401 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    11461 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    11521 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    11581 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    11641 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    11701 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    11761 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    11821 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    11881 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    11941 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    12001 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    12061 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    12121 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
    12181 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
    12241 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
    12301 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
    12361 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
    12421 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
    12481 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
    12541 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
    12601 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
    12661 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
    12721 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
    12781 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
    12841 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
    12901 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
    12961 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
    13021 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
    13081 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
    13141 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
    13201 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
    13261 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
    13321 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
    13381 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
    13441 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
    13501 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
    13561 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
    13621 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
    13681 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
    13741 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
    13801 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
    13861 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
    13921 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
    13981 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
    14041 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
    14101 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaata
//
