LOCUS       sequence-265907-       11964 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 11964)
  AUTHORS   Yeo NC, Chavez A, Lance-Byrne A, Chan Y, Menn D, Milanova D, Kuo CC,
            Guo X, Sharma S, Tung A, Cecchi RJ, Tuttle M, Pradhan S, Lim ET, 
            Davidsohn N, Ebrahimkhani MR, Collins JJ, Lewis NE, Kiani S, Church 
            GM
  TITLE     An enhanced CRISPR repressor for targeted mammalian gene regulation.
  JOURNAL   Nat Methods. 2018 Jul 16. pii: 10.1038/s41592-018-0048-5. doi: 
            10.1038/s41592-018-0048-5.
  PUBMED    30013045
REFERENCE   2  (bases 1 to 11964)
  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: "Nat 
            Methods. 2018 Jul 16. pii: 10.1038/s41592-018-0048-5. doi: 
            10.1038/s41592-018-0048-5."
COMMENT     Sequence Label: pB-CAGGS-dCas9-KRAB
FEATURES             Location/Qualifiers
     source          1..11964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1357..1736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1738..2015
                     /label=chicken beta-actin promoter
     intron          2016..3032
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2956..2978
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3040..3059
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3129..3138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3135..3161,3162..3164,3165..5648,5649..5654,5655..5720,
                     5721..5723,5724..7238)
                     /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             7251..7271
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7332..7526
                     /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(7581..7600)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7625..7673
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7905..8116
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8126..8135
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8132..8530
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8534..8655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8571..8590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8625..8644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9418..9452
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9943..9959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9943..9959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9956..9978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9967..9983
                     /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(9991..9997,9998..10015,10016..10021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10036..10057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10174..10191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10345..10933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10425..10444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11104..11895,11896..11964))
                     /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     11727..11746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccgaca ttgattattg actagttatt
     1381 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1441 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     1501 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     1561 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     1621 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1681 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     1741 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     1801 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
     1861 gcgcgccrgg sggggsgggg sggggsgrgg ggsggggsgg ggsgaggcgg agaggtgcgg
     1921 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     1981 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     2041 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     2101 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     2161 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     2221 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     2281 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttttg
     2341 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     2401 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     2461 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     2521 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     2581 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggrgggct
     2641 cgggggrggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     2701 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     2761 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     2821 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     2881 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     2941 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     3001 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     3061 tgtctcatca ttttggcaaa gaatctagat accctcgtaa aggtctagag ctagcggatc
     3121 gaacccttgc caccatggac aagaagtact ccattgggct cgctatcggc acaaacagcg
     3181 tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
     3241 gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg
     3301 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
     3361 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
     3421 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc
     3481 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
     3541 atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc
     3601 tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
     3661 cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt
     3721 tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc
     3781 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
     3841 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
     3901 tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg
     3961 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
     4021 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
     4081 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
     4141 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
     4201 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
     4261 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag
     4321 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
     4381 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
     4441 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
     4501 cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
     4561 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
     4621 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
     4681 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
     4741 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
     4801 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
     4861 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
     4921 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
     4981 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
     5041 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat
     5101 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg
     5161 gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt
     5221 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
     5281 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
     5341 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
     5401 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
     5461 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
     5521 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
     5581 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact
     5641 acgacgtggc tgctatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag
     5701 tgttgacaag atccgataaa gctagaggga agagtgataa cgtcccctca gaagaagttg
     5761 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
     5821 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
     5881 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
     5941 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
     6001 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
     6061 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
     6121 cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt
     6181 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
     6241 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
     6301 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
     6361 agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta
     6421 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
     6481 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
     6541 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
     6601 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa
     6661 aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca
     6721 ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
     6781 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
     6841 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
     6901 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat
     6961 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
     7021 agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga
     7081 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt
     7141 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
     7201 atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac cccaagaaga
     7261 agaggaaggt gagtggtgga ggaagtggcg ggtcagggtc gatggacgcg aaatcactta
     7321 cggcatggtc gagaacactg gttacgttca aggacgtgtt tgtggacttt acacgtgagg
     7381 agtggaaatt gctggatact gcgcaacaaa ttgtgtatcg aaatgtcatg cttgagaatt
     7441 acaagaacct cgtcagtctc ggataccagt tgacgaaacc ggatgtgatc cttaggctcg
     7501 aaaaggggga agaaccttgg ctggtataga agaattcgat ccctaccggt tagtaatgag
     7561 tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     7621 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     7681 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     7741 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7801 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgaa atcggatccg
     7861 cggccgcact gattctagct gcagaggctc cggtgcccgt cagtgggcag agcgcacatc
     7921 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     7981 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     8041 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     8101 tgccgccaga acacaggatc ctcgagccac catggccaag cctttgtctc aagaagaatc
     8161 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     8221 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     8281 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     8341 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     8401 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     8461 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     8521 ggagggctaa gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     8581 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     8641 catcaatgta tcttatcatg tctggaattg actcaaatga tgtcaattag tctatcagaa
     8701 gctcatctgg tctcccttcc gggggacaag acatccctgt ttaatattta aacagcagtg
     8761 ttcccaaact gggttcttat atcccttgct ctggtcaacc aggttgcagg gtttcctgtc
     8821 ctcacaggaa cgaagtccct aaagaaacag tggcagccag gtttagcccc ggaattgact
     8881 ggattccttt tttagggccc attggtatgg ctttttcccc gtatcccccc aggtgtctgc
     8941 aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg
     9001 cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg
     9061 gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg
     9121 ggctttgggg gggggctgtc cctgatatct ataacaagaa aatatatata taataagtta
     9181 tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc
     9241 acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg
     9301 acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct
     9361 aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat
     9421 gcgtcaattt tacgcagact atctttctag ggttaatcta gctgcatcag gatcatatcg
     9481 tcgggtcttt tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg
     9541 aagaagcccg tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg
     9601 actgctgctg cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg
     9661 tggccatgca cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt
     9721 cgcggctttt ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca
     9781 ataccggcga cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc
     9841 tgggatatta cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga
     9901 taccccgtga gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     9961 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    10021 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    10081 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
    10141 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    10201 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    10261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    10321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    10381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    10441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    10501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    10561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    10621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    10681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    10741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
    10801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    10861 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    10921 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    10981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
    11041 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
    11101 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
    11161 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
    11221 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
    11281 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    11341 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    11401 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    11461 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    11521 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    11581 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    11641 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    11701 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    11761 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    11821 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    11881 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    11941 gacacggaaa tgttgaatac tcat
//
