LOCUS       sequence-354076-       12115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12115)
  AUTHORS   Nghi Dang , Alissa Lance-Byrne, Angela Tung, Xiaoge Guo, Ryan J 
            Cecchi, Joanna Buchthal, Alejandro Chavez, Nan Cher Yeo
  TITLE     Generation and application of a versatile CRISPR toolkit for 
            mammalian cell engineering
  JOURNAL   Synthetic Biology, Volume 6, Issue 1, 2021
REFERENCE   2  (bases 1 to 12115)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synthetic 
            Biology, Volume 6, Issue 1, 2021"
COMMENT     Sequence Label: NAX_CAGGS_CAS9_m4_KRAB_Blasto
FEATURES             Location/Qualifiers
     source          1..12115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          145..1161
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1085..1107
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1169..1188
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1258..1267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1264..1290,1291..1293,1294..3777,3778..3783,3784..3849,
                     3850..3852,3853..5367)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5380..5400
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5461..5655
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     primer_bind     complement(5710..5729)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5754..5802
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6034..6245
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6255..6264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6261..6659
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6663..6784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6700..6719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6754..6773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7547..7581
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8072..8088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8072..8088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8085..8107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8096..8112
                     /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(8120..8126,8127..8144,8145..8150))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8165..8186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8303..8320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8474..9062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8554..8573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9233..10024,10025..10093))
                     /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     9856..9875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10094..10198)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10224..10679)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10361..10382)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10573..10592
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10806..10828
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10820..10837
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10821..10837
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10967..11029
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11450..11829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
       61 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      121 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      181 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      241 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      301 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
      361 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
      421 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttttgc gctccgcagt
      481 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
      541 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
      601 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
      661 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
      721 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
      781 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
      841 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
      901 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
      961 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     1021 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1081 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1141 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1201 tttggcaaag aatctagata ccctcgtaaa ggtctagagc tagcggatcg aacccttgcc
     1261 accatggaca agaagtactc cattgggctc gctatcggca caaacagcgt cggctgggcc
     1321 gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat
     1381 cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc
     1441 gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc
     1501 tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat
     1561 aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt
     1621 ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg
     1681 aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg
     1741 catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc
     1801 gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac
     1861 ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc
     1921 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt
     1981 aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc
     2041 gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg
     2101 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc
     2161 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct
     2221 agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc
     2281 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac
     2341 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc
     2401 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg
     2461 cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg
     2521 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag
     2581 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat
     2641 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag
     2701 gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat
     2761 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca
     2821 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc
     2881 ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt
     2941 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa
     3001 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa
     3061 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt
     3121 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac
     3181 gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg
     3241 gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc
     3301 ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat
     3361 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt
     3421 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag
     3481 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc
     3541 gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa
     3601 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac
     3661 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc
     3721 agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggct
     3781 gctatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga
     3841 tccgataaag ctagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg
     3901 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat
     3961 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg
     4021 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg
     4081 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag
     4141 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac
     4201 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa
     4261 aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg
     4321 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac
     4381 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag
     4441 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat
     4501 ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa
     4561 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg
     4621 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc
     4681 gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc
     4741 gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc
     4801 gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc
     4861 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag
     4921 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc
     4981 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg
     5041 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga
     5101 gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat
     5161 aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc
     5221 gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca
     5281 aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga
     5341 atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg
     5401 agtggtggag gaagtggcgg gtcagggtcg atggacgcga aatcacttac ggcatggtcg
     5461 agaacactgg ttacgttcaa ggacgtgttt gtggacttta cacgtgagga gtggaaattg
     5521 ctggatactg cgcaacaaat tgtgtatcga aatgtcatgc ttgagaatta caagaacctc
     5581 gtcagtctcg gataccagtt gacgaaaccg gatgtgatcc ttaggctcga aaagggggaa
     5641 gaaccttggc tggtatagaa gaattcgatc cctaccggtt agtaatgagt ttaaacgggg
     5701 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5761 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5821 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5881 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5941 acgtcggggc ggcaggccct gccatagcag atctgcgaaa tcggatccgc ggccgcactg
     6001 attctagctg cagaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     6061 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
     6121 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     6181 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     6241 cacaggatcc tcgagccacc atggccaagc ctttgtctca agaagaatcc accctcattg
     6301 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     6361 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     6421 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     6481 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     6541 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     6601 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag
     6661 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6721 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6781 cttatcatgt ctggaattga ctcaaatgat gtcaattagt ctatcagaag ctcatctggt
     6841 ctcccttccg ggggacaaga catccctgtt taatatttaa acagcagtgt tcccaaactg
     6901 ggttcttata tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac
     6961 gaagtcccta aagaaacagt ggcagccagg tttagccccg gaattgactg gattcctttt
     7021 ttagggccca ttggtatggc tttttccccg tatcccccca ggtgtctgca ggctcaaaga
     7081 gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc
     7141 cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc
     7201 ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg
     7261 ggggctgtcc ctgatatcta taacaagaaa atatatatat aataagttat cacgtaagta
     7321 gaacatgaaa taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat
     7381 aatcatgcgt cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc
     7441 attgacaagc acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc
     7501 gacggattcg cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt
     7561 acgcagacta tctttctagg gttaatctag ctgcatcagg atcatatcgt cgggtctttt
     7621 ttccggctca gtcatcgccc aagctggcgc tatctgggca tcggggagga agaagcccgt
     7681 gccttttccc gcgaggttga agcggcatgg aaagagtttg ccgaggatga ctgctgctgc
     7741 attgacgttg agcgaaaacg cacgtttacc atgatgattc gggaaggtgt ggccatgcac
     7801 gcctttaacg gtgaactgtt cgttcaggcc acctgggata ccagttcgtc gcggcttttc
     7861 cggacacagt tccggatggt cagcccgaag cgcatcagca acccgaacaa taccggcgac
     7921 agccggaact gccgtgccgg tgtgcagatt aatgacagcg gtgcggcgct gggatattac
     7981 gtcagcgagg acgggtatcc tggctggatg ccgcagaaat ggacatggat accccgtgag
     8041 ttacccggcg ggcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     8101 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     8161 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     8221 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     8281 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     8341 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     8401 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     8461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8881 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     9181 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9301 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9361 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9421 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9481 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9541 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9601 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9661 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9721 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9781 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9841 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9901 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9961 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10021 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    10081 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    10141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    10201 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
    10261 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
    10321 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
    10381 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
    10441 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
    10501 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
    10561 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
    10621 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
    10681 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
    10741 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
    10801 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt
    10861 ttttgaaccc gtggaggacg ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga
    10921 gcagatgaag atgctcgaca cgctgcagaa cacgcagcta gattaaccct agaaagataa
    10981 tcatattgtg acgtacgtta aagataatca tgtgtaaaat tgacgcatgt gttttatcgg
    11041 tctgtatatc gaggtttatt tattaatttg aatagatatt aagttttatt atatttacac
    11101 ttacatacta ataataaatt caacaaacaa tttatttatg tttatttatt tattaaaaaa
    11161 aacaaaaact caaaatttct tctataaagt aacaaaactt ttatgaggga cagccccccc
    11221 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
    11281 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
    11341 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
    11401 ttgagcctgc agacacctgg ggggatacgg ggaaaaggcc tccacggccg acattgatta
    11461 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
    11521 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
    11581 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
    11641 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
    11701 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
    11761 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
    11821 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
    11881 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
    11941 gggggggggg gggcgcgccc gcgggggggg gggggggggg gggggggggg gggggggggg
    12001 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
    12061 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg ggggg
//
