LOCUS       sequence-354085-       11823 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 11823)
  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 11823)
  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_Hygro
FEATURES             Location/Qualifiers
     source          1..11823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     321..343
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     405..424
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      494..503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(500..526,527..529,530..3013,3014..3019,3020..3085,
                     3086..3088,3089..4603)
                     /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             4616..4636
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4697..4891
                     /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(4946..4965)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4990..5038
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5270..5481
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5491..5500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5512..6531
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGTGTVGRTQIARRGAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     polyA_signal    6535..6656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6572..6591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6626..6645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7419..7453
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7944..7960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7944..7960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7957..7979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7968..7984
                     /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(7992..7998,7999..8016,8017..8022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8037..8058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8175..8192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8346..8934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8426..8445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9105..9896,9897..9965))
                     /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     9728..9747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9966..10070)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10096..10551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10233..10254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10445..10464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10678..10700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10692..10709
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10693..10709
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10839..10901
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11322..11701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
       61 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      121 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
      181 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
      241 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
      301 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
      361 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
      421 tgtgctgtct catcattttg gcaaagaatc tagataccct cgtaaaggtc tagagctagc
      481 ggatcgaacc cttgccacca tggacaagaa gtactccatt gggctcgcta tcggcacaaa
      541 cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt
      601 tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga
      661 ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg
      721 cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga
      781 tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga
      841 gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac
      901 catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat
      961 ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct
     1021 gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca
     1081 gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc
     1141 taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa
     1201 gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc
     1261 taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga
     1321 tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa
     1381 gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa
     1441 agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt
     1501 gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca
     1561 gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa
     1621 atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa
     1681 cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat
     1741 tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa
     1801 agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc
     1861 cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac
     1921 tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag
     1981 gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct
     2041 gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat
     2101 gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa
     2161 gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg
     2221 tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta
     2281 tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga
     2341 cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga
     2401 acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg
     2461 ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc gagacaagca
     2521 gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc ggaacttcat
     2581 gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag cacaagtttc
     2641 tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc cagctatcaa
     2701 aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca
     2761 taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc agaagggaca
     2821 gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac tggggtccca
     2881 aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc tctacctgta
     2941 ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca atcggctctc
     3001 cgactacgac gtggctgcta tcgtgcccca gtcttttctc aaagatgatt ctattgataa
     3061 taaagtgttg acaagatccg ataaagctag agggaagagt gataacgtcc cctcagaaga
     3121 agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac tgatcacaca
     3181 acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt tggataaagc
     3241 cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg tggcccaaat
     3301 tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc gagaggtgaa
     3361 agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc agttttataa
     3421 ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg cagtggtagg
     3481 cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg gagactataa
     3541 agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca aggccaccgc
     3601 taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta cactggccaa
     3661 tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag aaatcgtgtg
     3721 ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc aggtgaacat
     3781 cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc tcccgaaaag
     3841 gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat acggcggatt
     3901 cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga aagggaagtc
     3961 taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc gatcaagctt
     4021 cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca aaaaagacct
     4081 catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga aacgaatgct
     4141 cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta aatacgttaa
     4201 tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag ataatgagca
     4261 gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg agcaaataag
     4321 cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc tttctgctta
     4381 caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc acttgtttac
     4441 tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca tagacagaaa
     4501 gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt caattacggg
     4561 gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg ctgaccccaa
     4621 gaagaagagg aaggtgagtg gtggaggaag tggcgggtca gggtcgatgg acgcgaaatc
     4681 acttacggca tggtcgagaa cactggttac gttcaaggac gtgtttgtgg actttacacg
     4741 tgaggagtgg aaattgctgg atactgcgca acaaattgtg tatcgaaatg tcatgcttga
     4801 gaattacaag aacctcgtca gtctcggata ccagttgacg aaaccggatg tgatccttag
     4861 gctcgaaaag ggggaagaac cttggctggt atagaagaat tcgatcccta ccggttagta
     4921 atgagtttaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4981 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     5041 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5101 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5161 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgaaatcgg
     5221 atccgcggcc acactgattc tagctgcaga ggctccggtg cccgtcagtg ggcagagcgc
     5281 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
     5341 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     5401 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct tttccgcaac
     5461 gggtttgccg ccagaacaca ggatcctcga gccaccatgg atagatccgg aaagcctgaa
     5521 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ttccgacctg
     5581 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     5641 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     5701 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     5761 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     5821 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     5881 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     5941 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     6001 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     6061 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     6121 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     6181 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     6241 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     6301 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     6361 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat caggaaccgg gactgtcggg
     6421 cgtacacaaa tcgcccgcag aggcgcggcc gtctggaccg atggctgtgt agaagtactc
     6481 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata ggttaacttg
     6541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6661 gtctggaatt gactcaaatg atgtcaatta gtctatcaga agctcatctg gtctcccttc
     6721 cgggggacaa gacatccctg tttaatattt aaacagcagt gttcccaaac tgggttctta
     6781 tatcccttgc tctggtcaac caggttgcag ggtttcctgt cctcacagga acgaagtccc
     6841 taaagaaaca gtggcagcca ggtttagccc cggaattgac tggattcctt ttttagggcc
     6901 cattggtatg gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     6961 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     7021 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     7081 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     7141 ccctgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
     7201 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
     7261 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
     7321 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
     7381 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac
     7441 tatctttcta gggttaatct agctgcatca ggatcatatc gtcgggtctt ttttccggct
     7501 cagtcatcgc ccaagctggc gctatctggg catcggggag gaagaagccc gtgccttttc
     7561 ccgcgaggtt gaagcggcat ggaaagagtt tgccgaggat gactgctgct gcattgacgt
     7621 tgagcgaaaa cgcacgttta ccatgatgat tcgggaaggt gtggccatgc acgcctttaa
     7681 cggtgaactg ttcgttcagg ccacctggga taccagttcg tcgcggcttt tccggacaca
     7741 gttccggatg gtcagcccga agcgcatcag caacccgaac aataccggcg acagccggaa
     7801 ctgccgtgcc ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt acgtcagcga
     7861 ggacgggtat cctggctgga tgccgcagaa atggacatgg ataccccgtg agttacccgg
     7921 cgggcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     7981 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     8041 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     8101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     8161 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     8221 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     8281 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     8341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     8401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     8461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     8521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     8581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     8641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     8701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     8761 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     8821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     8881 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8941 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     9001 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9061 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9181 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     9241 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     9301 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     9361 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     9421 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     9481 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     9541 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     9601 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     9661 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9721 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9781 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9841 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9901 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9961 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    10021 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    10081 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
    10141 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
    10201 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
    10261 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
    10321 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
    10381 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
    10441 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
    10501 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
    10561 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
    10621 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
    10681 agtcacgacg ttgtaaaacg acggccagtg agcgcgcctc gttcattcac gtttttgaac
    10741 ccgtggagga cgggcagact cgcggtgcaa atgtgtttta cagcgtgatg gagcagatga
    10801 agatgctcga cacgctgcag aacacgcagc tagattaacc ctagaaagat aatcatattg
    10861 tgacgtacgt taaagataat catgtgtaaa attgacgcat gtgttttatc ggtctgtata
    10921 tcgaggttta tttattaatt tgaatagata ttaagtttta ttatatttac acttacatac
    10981 taataataaa ttcaacaaac aatttattta tgtttattta tttattaaaa aaaacaaaaa
    11041 ctcaaaattt cttctataaa gtaacaaaac ttttatgagg gacagccccc ccccaaagcc
    11101 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
    11161 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
    11221 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
    11281 gcagacacct ggggggatac ggggaaaagg cctccacggc cgacattgat tattgactag
    11341 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
    11401 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    11461 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
    11521 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
    11581 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
    11641 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
    11701 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
    11761 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
    11821 ggg
//
