LOCUS       sequence-271169-       12008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 in a transposon array under a Tet inducible promoter. For 
            making inducible Cas9 cell lines. Neo/G418 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12008)
  AUTHORS   McKinley KL, Sekulic N, Guo LY, Tsinman T, Black BE, Cheeseman IM
  TITLE     The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork
            of Interactions at the Centromere-Kinetochore Interface.
  JOURNAL   Mol Cell. 2015 Dec 17;60(6):886-98. doi: 
            10.1016/j.molcel.2015.10.027. Epub 2015 Nov 19.
  PUBMED    26698661
REFERENCE   2  (bases 1 to 12008)
  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.2015.10.027"; journalName: "Mol Cell"; date: 
            "2015-12-17- 17"; volume: "60"; issue: "6"; pages: "886-98"
COMMENT     Sequence Label: HP138-neo
FEATURES             Location/Qualifiers
     source          1..12008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        413..791
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    421..439
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    457..475
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    493..511
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    529..547
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    565..583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    601..619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    637..655
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     700..724
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      818..827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             824..4927
                     /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             824..4927
                     /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             824..4927
                     /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             4940..4960
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5015..5034)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5059..5107
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5338..5549
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5562..5830
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             5864..6607
                     /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             6608..6661
                     /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"
     primer_bind     complement(6662..6681)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             6710..7513
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6773..6792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7383..7402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7606..7727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7643..7662)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7697..7716
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8490..8524
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9015..9031)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9015..9031)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9028..9050)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9039..9055
                     /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(9063..9069,9070..9087,9088..9093))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9108..9129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9246..9263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9417..10005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9497..9516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10176..10967,10968..11036))
                     /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     10799..10818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11037..11141)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11167..11622)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11304..11325)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11516..11535
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11749..11771
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11763..11780
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11764..11780
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11910..11972
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 atttgaatag atattaagtt ttattatatt tacacttaca tactaataat aaattcaaca
       61 aacaatttat ttatgtttat ttatttatta aaaaaaacaa aaactcaaaa tttcttctat
      121 aaagtaacaa aacttttatg agggacagcc cccccccaaa gcccccaggg atgtaattac
      181 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
      241 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
      301 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
      361 tacggggaaa aggcctccac ggccactagt ctttcgtctt caagaattcc tcgagtttac
      421 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
      481 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
      541 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
      601 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
      661 taagctttag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      721 gatcgcctgg agcaattcca caacactttt gtcttatacc aactttccgt accacttcct
      781 accctcgtaa aggtctagag ctagcgaatt cgaatttgcc accatggaca agaagtactc
      841 cattgggctc gatatcggca caaacagcgt cggctgggcc gtcattacgg acgagtacaa
      901 ggtgccgagc aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa
      961 cctcattggc gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag
     1021 aacagcacgg cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt
     1081 tagtaatgag atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt
     1141 ggtggaggag gataaaaagc acgagcgcca cccaatcttt ggcaatatcg tggacgaggt
     1201 ggcgtaccat gaaaagtacc caaccatata tcatctgagg aagaagcttg tagacagtac
     1261 tgataaggct gacttgcggt tgatctatct cgcgctggcg catatgatca aatttcgggg
     1321 acacttcctc atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat
     1381 ccaactggtt cagacttaca atcagctttt cgaagagaac ccgatcaacg catccggagt
     1441 tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat
     1501 cgcacagctc cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact
     1561 cgggctgacc cccaacttta aatctaactt cgacctggcc gaagatgcca agcttcaact
     1621 gagcaaagac acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta
     1681 cgcagacctt tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct
     1741 gcgagtgaac acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga
     1801 tgagcaccac caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa
     1861 gtacaaggaa attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg
     1921 agcaagccag gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac
     1981 cgaggagctg ctggtaaagc ttaacagaga agatctgttg cgcaaacagc gcactttcga
     2041 caatggaagc atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca
     2101 agaggatttc tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt
     2161 tcggataccc tactatgtag gccccctcgc ccggggaaat tccagattcg cgtggatgac
     2221 tcgcaaatca gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc
     2281 ctctgcccag tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa
     2341 ggtgcttcct aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa
     2401 ggtcaaatac gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa
     2461 agctatcgtg gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga
     2521 agactatttc aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg
     2581 cttcaacgca tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt
     2641 cctggacaat gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt
     2701 tgaagatagg gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa
     2761 agtcatgaaa cagctcaaga ggcgccgata tacaggatgg gggcggctgt caagaaaact
     2821 gatcaatggg atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga
     2881 tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga
     2941 ggacatccag aaagcacaag tttctggcca gggggacagt cttcacgagc acatcgctaa
     3001 tcttgcaggt agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga
     3061 actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc gttatcgaga tggcccgaga
     3121 gaaccaaact acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga
     3181 gggtataaaa gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct
     3241 tcagaatgag aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca
     3301 ggaactggac atcaatcggc tctccgacta cgacgtggat catatcgtgc cccagtcttt
     3361 tctcaaagat gattctattg ataataaagt gttgacaaga tccgataaaa atagagggaa
     3421 gagtgataac gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct
     3481 gctgaacgcc aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg
     3541 tggcctgtct gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca
     3601 gatcaccaag cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa
     3661 tgacaaactg attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt
     3721 cagaaaggac tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga
     3781 tgcctacctg aatgcagtgg taggcactgc acttatcaaa aaatatccca agcttgaatc
     3841 tgaatttgtt tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga
     3901 gcaggaaata ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt
     3961 caagaccgag attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa
     4021 cggagaaaca ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccggaaggt
     4081 cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc
     4141 caaggaaagt atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg
     4201 ggaccccaag aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt
     4261 ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat
     4321 cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg
     4381 atataaagag gtcaaaaaag acctcatcat taagcttccc aagtactctc tctttgagct
     4441 tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct
     4501 ggcactgccc tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa
     4561 agggtctccc gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct
     4621 tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa
     4681 cctcgataag gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc
     4741 agaaaacatt atccacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta
     4801 cttcgacacc accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac
     4861 actgattcat cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg
     4921 tggagacagc agggctgacc ccaagaagaa gaggaaggtg tgaaagggtt cgatccctac
     4981 cggttagtaa tgagtttaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     5041 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     5101 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5161 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5221 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     5281 cgaaatcgga tccagctgaa tctaagtgga tctgcgatcg ctccggtgcc cgtcagtggg
     5341 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg
     5401 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     5461 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     5521 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt
     5581 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc
     5641 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt
     5701 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca
     5761 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc
     5821 gttacagatc taagctgtga ccggcgccta ctctagagcc gccatgtcta gactggacaa
     5881 gagcaaagtc ataaactctg ctctggaatt actcaatgga gtcggtatcg aaggcctgac
     5941 gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa
     6001 gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca
     6061 ctcctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata
     6121 ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac
     6181 agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc
     6241 cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt
     6301 ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat
     6361 gcccccactt ctgaaacaag caattgagct gttcgaccgg cagggagccg aacctgcctt
     6421 ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg
     6481 cgggccgacc gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga
     6541 cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct
     6601 ccccggggag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     6661 tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtca tgggatcggc
     6721 cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     6781 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     6841 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     6901 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     6961 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     7021 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     7081 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     7141 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     7201 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     7261 cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     7321 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     7381 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     7441 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     7501 cgagttcttc tgagacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     7561 gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gggtaaactt gtttattgca
     7621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     7681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggaat
     7741 tgactcaaat gatgtcaatt agtctatcag aagctcatct ggtctccctt ccgggggaca
     7801 agacatccct gtttaatatt taaacagcag tgttcccaaa ctgggttctt atatcccttg
     7861 ctctggtcaa ccaggttgca gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac
     7921 agtggcagcc aggtttagcc ccggaattga ctggattcct tttttagggc ccattggtat
     7981 ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca
     8041 gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct
     8101 cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc
     8161 tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg tccctgatat
     8221 ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat
     8281 ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg
     8341 actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc
     8401 acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt
     8461 tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct
     8521 agggttaatc tagctgcatc aggatcatat cgtcgggtct tttttccggc tcagtcatcg
     8581 cccaagctgg cgctatctgg gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt
     8641 tgaagcggca tggaaagagt ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa
     8701 acgcacgttt accatgatga ttcgggaagg tgtggccatg cacgccttta acggtgaact
     8761 gttcgttcag gccacctggg ataccagttc gtcgcggctt ttccggacac agttccggat
     8821 ggtcagcccg aagcgcatca gcaacccgaa caataccggc gacagccgga actgccgtgc
     8881 cggtgtgcag attaatgaca gcggtgcggc gctgggatat tacgtcagcg aggacgggta
     8941 tcctggctgg atgccgcaga aatggacatg gataccccgt gagttacccg gcgggcgcgc
     9001 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     9061 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     9121 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     9181 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     9241 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     9301 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     9361 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     9421 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     9481 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     9541 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     9601 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     9661 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     9721 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     9781 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     9841 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9901 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9961 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    10021 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    10081 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    10141 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    10201 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    10261 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    10321 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    10381 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    10441 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    10501 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    10561 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    10621 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    10681 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    10741 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    10801 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    10861 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    10921 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    10981 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    11041 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    11101 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    11161 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
    11221 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
    11281 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
    11341 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
    11401 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
    11461 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
    11521 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
    11581 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
    11641 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
    11701 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
    11761 gttgtaaaac gacggccagt gagcgcgcct cgttcattca cgtttttgaa cccgtggagg
    11821 acgggcagac tcgcggtgca aatgtgtttt acagcgtgat ggagcagatg aagatgctcg
    11881 acacgctgca gaacacgcag ctagattaac cctagaaaga taatcatatt gtgacgtacg
    11941 ttaaagataa tcatgtgtaa aattgacgca tgtgttttat cggtctgtat atcgaggttt
    12001 atttatta
//
