LOCUS       sequence-271170-       11696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 in a transposon array under a Tet inducible promoter. For 
            making inducible Cas9 cell lines. Puro selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11696)
  AUTHORS   McKinley KL, Cheeseman IM
  TITLE     Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the
            Diversity of p53-Dependent Responses to Cell-Cycle Defects.
  JOURNAL   Dev Cell. 2017 Feb 27;40(4):405-420.e2. doi: 
            10.1016/j.devcel.2017.01.012. Epub 2017 Feb 16.
  PUBMED    28216383
REFERENCE   2  (bases 1 to 11696)
  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.devcel.2017.01.012"; journalName: "Dev Cell"; date: 
            "2017-02-27- 27"; volume: "40"; issue: "4"; pages: "405-420.e2"
COMMENT     Sequence Label: HP138-puro
FEATURES             Location/Qualifiers
     source          1..11696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        323..701
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    331..349
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    367..385
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    403..421
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    439..457
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    475..493
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    511..529
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    547..565
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     610..634
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      728..737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             734..4837
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             734..4837
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             734..4837
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4850..4870
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4925..4944)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4969..5017
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5248..5459
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5472..5740
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             5774..6517
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             6518..6571
                     /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             6572..7171
                     /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(6572..6591)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7068..7088
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7204..7325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7241..7260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7295..7314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8088..8122
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8613..8629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8613..8629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8626..8648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8637..8653
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8661..8667,8668..8685,8686..8691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8706..8727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8844..8861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9015..9603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9095..9114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9774..10565,10566..10634))
                     /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     10397..10416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10635..10739)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10765..11220)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10902..10923)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11114..11133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11347..11369
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11361..11378
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11362..11378
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11508..11570
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttatg agggacagcc
       61 cccccccaaa gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag
      121 ccgcccgggg ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg
      181 gggacagccc gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct
      241 gctctttgag cctgcagaca cctgggggga tacggggaaa aggcctccac ggccactagt
      301 ctttcgtctt caagaattcc tcgagtttac tccctatcag tgatagagaa cgtatgaaga
      361 gtttactccc tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag
      421 aacgtataag gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta
      481 tcagtgatag agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca
      541 gtttactccc tatcagtgat agagaacgta taagctttag gcgtgtacgg tgggcgccta
      601 taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt
      661 gtcttatacc aactttccgt accacttcct accctcgtaa aggtctagag ctagcgaatt
      721 cgaatttgcc accatggaca agaagtactc cattgggctc gatatcggca caaacagcgt
      781 cggctgggcc gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg
      841 caataccgat cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg
      901 ggagacggcc gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa
      961 gaatcggatc tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc
     1021 tttcttccat aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca
     1081 cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata
     1141 tcatctgagg aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct
     1201 cgcgctggcg catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc
     1261 agacaacagc gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt
     1321 cgaagagaac ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct
     1381 gtccaaatcc cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg
     1441 cctgtttggt aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt
     1501 cgacctggcc gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga
     1561 caatctgctg gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct
     1621 gtcagacgcc attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc
     1681 gctgagcgct agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa
     1741 ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa
     1801 aaatggctac gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat
     1861 taagcccatc ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga
     1921 agatctgttg cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct
     1981 gggcgaactg cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa
     2041 cagggaaaag attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc
     2101 ccggggaaat tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg
     2161 gaacttcgag gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac
     2221 taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga
     2281 gtacttcaca gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa
     2341 gccagcattc ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa
     2401 ccggaaagtt accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga
     2461 ctctgttgaa atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga
     2521 tctcctgaaa atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct
     2581 tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt
     2641 gaaaacttac gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata
     2701 tacaggatgg gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg
     2761 aaagacaatc ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt
     2821 gatccatgat gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca
     2881 gggggacagt cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg
     2941 aatactgcag accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc
     3001 cgagaatatc gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa
     3061 cagtagggaa aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct
     3121 taaggaacac ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct
     3181 gcagaacggc agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta
     3241 cgacgtggat catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt
     3301 gttgacaaga tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt
     3361 caagaaaatg aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa
     3421 gttcgataat ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt
     3481 catcaaaagg cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga
     3541 ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat
     3601 tactctgaag tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag
     3661 agagatcaac aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc
     3721 acttatcaaa aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta
     3781 cgatgttagg aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta
     3841 cttcttttac agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga
     3901 gattcggaag cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa
     3961 gggtagggat ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa
     4021 aaagaccgaa gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag
     4081 cgacaagctg atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc
     4141 tcctacagtc gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa
     4201 actcaaaagc gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa
     4261 aaaccccatc gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat
     4321 taagcttccc aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag
     4381 tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt
     4441 gtatctggcc agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca
     4501 gctgttcgtg gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt
     4561 ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa
     4621 gcacagggat aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac
     4681 caacttgggc gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta
     4741 cacctctaca aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta
     4801 tgaaacaaga atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa
     4861 gaggaaggtg tgaaagggtt cgatccctac cggttagtaa tgagtttaaa cgggggaggc
     4921 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4981 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5041 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5101 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5161 ggggcggcag gccctgccat agcagatctg cgaaatcgga tccagctgaa tctaagtgga
     5221 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     5281 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact
     5341 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     5401 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc
     5461 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
     5521 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
     5581 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
     5641 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
     5701 acgtctttgt ttcgttttct gttctgcgcc gttacagatc taagctgtga ccggcgccta
     5761 ctctagagcc gccatgtcta gactggacaa gagcaaagtc ataaactctg ctctggaatt
     5821 actcaatgga gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt
     5881 tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc
     5941 aatcgagatg ctggacaggc atcataccca ctcctgcccc ctggaaggcg agtcatggca
     6001 agactttctg cggaacaacg ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg
     6061 ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc tggaaaatca
     6121 gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg ctctgtccgc
     6181 cgtgggccac tttacactgg gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga
     6241 ggaaagagag acacctacca ccgattctat gcccccactt ctgaaacaag caattgagct
     6301 gttcgaccgg cagggagccg aacctgcctt ccttttcggc ctggaactaa tcatatgtgg
     6361 cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg acgattttga
     6421 cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc tgcctgctga
     6481 cgctcttgac gattttgacc ttgacatgct ccccggggag ggcagaggaa gtcttctaac
     6541 atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct
     6601 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     6661 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     6721 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     6781 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     6841 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     6901 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     6961 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     7021 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     7081 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     7141 ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtgg
     7201 gtaaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7321 tcttatcatg tctggaattg actcaaatga tgtcaattag tctatcagaa gctcatctgg
     7381 tctcccttcc gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact
     7441 gggttcttat atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa
     7501 cgaagtccct aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt
     7561 tttagggccc attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
     7621 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
     7681 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
     7741 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
     7801 gggggctgtc cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt
     7861 agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga
     7921 taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg
     7981 cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag
     8041 cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt
     8101 tacgcagact atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt
     8161 tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg
     8221 tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg
     8281 cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca
     8341 cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt
     8401 ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga
     8461 cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta
     8521 cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga
     8581 gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     8641 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     8701 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     8761 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     8821 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     8881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     8941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     9001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     9061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     9121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     9181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     9241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     9301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     9361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     9421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     9481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     9541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     9601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     9661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     9721 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     9781 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     9841 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     9901 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     9961 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    10021 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    10081 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    10141 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    10201 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    10261 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    10321 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    10381 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    10441 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    10501 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    10561 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    10621 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    10681 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    10741 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
    10801 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
    10861 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
    10921 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
    10981 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
    11041 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
    11101 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
    11161 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
    11221 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
    11281 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
    11341 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcctcg ttcattcacg
    11401 tttttgaacc cgtggaggac gggcagactc gcggtgcaaa tgtgttttac agcgtgatgg
    11461 agcagatgaa gatgctcgac acgctgcaga acacgcagct agattaaccc tagaaagata
    11521 atcatattgt gacgtacgtt aaagataatc atgtgtaaaa ttgacgcatg tgttttatcg
    11581 gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca
    11641 cttacatact aataataaat tcaacaaaca atttatttat gtttatttat ttatta
//
