LOCUS       sequence-354081-       12274 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 12274)
  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 12274)
  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_EBFP2
FEATURES             Location/Qualifiers
     source          1..12274
                     /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             join(6261..6263,6264..6266,6267..6980)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(6306..6327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    6984..7105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7021..7040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7075..7094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7868..7902
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8393..8409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8393..8409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8406..8428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8417..8433
                     /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(8441..8447,8448..8465,8466..8471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8486..8507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8624..8641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8795..9383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8875..8894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9554..10345,10346..10414))
                     /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     10177..10196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10415..10519)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10545..11000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10682..10703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10894..10913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11127..11149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11141..11158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11142..11158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11288..11350
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11771..12150
                     /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 atggtgagca agggcgagga gctgttcacc ggggtggtgc
     6301 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg
     6361 gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     6421 tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc
     6481 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     6541 tccaggagcg caccatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga
     6601 agttcgaggg cgacacccta gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg
     6661 acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaac atctatatca
     6721 tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacgtggagg
     6781 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     6841 tgctgctgcc cgacagccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg
     6901 agaagcgcga tcacatggtc ctgctggagt tccgcaccgc cgccgggatc actctcggca
     6961 tggacgagct gtacaagtaa gttaacttgt ttattgcagc ttataatggt tacaaataaa
     7021 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7081 tgtccaaact catcaatgta tcttatcatg tctggaattg actcaaatga tgtcaattag
     7141 tctatcagaa gctcatctgg tctcccttcc gggggacaag acatccctgt ttaatattta
     7201 aacagcagtg ttcccaaact gggttcttat atcccttgct ctggtcaacc aggttgcagg
     7261 gtttcctgtc ctcacaggaa cgaagtccct aaagaaacag tggcagccag gtttagcccc
     7321 ggaattgact ggattccttt tttagggccc attggtatgg ctttttcccc gtatcccccc
     7381 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac
     7441 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg
     7501 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta
     7561 catccctggg ggctttgggg gggggctgtc cctgatatct ataacaagaa aatatatata
     7621 taataagtta tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat
     7681 cttaaaagtc acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc
     7741 aaaatcagtg acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact
     7801 gagatgtcct aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc
     7861 aagaatgcat gcgtcaattt tacgcagact atctttctag ggttaatcta gctgcatcag
     7921 gatcatatcg tcgggtcttt tttccggctc agtcatcgcc caagctggcg ctatctgggc
     7981 atcggggagg aagaagcccg tgccttttcc cgcgaggttg aagcggcatg gaaagagttt
     8041 gccgaggatg actgctgctg cattgacgtt gagcgaaaac gcacgtttac catgatgatt
     8101 cgggaaggtg tggccatgca cgcctttaac ggtgaactgt tcgttcaggc cacctgggat
     8161 accagttcgt cgcggctttt ccggacacag ttccggatgg tcagcccgaa gcgcatcagc
     8221 aacccgaaca ataccggcga cagccggaac tgccgtgccg gtgtgcagat taatgacagc
     8281 ggtgcggcgc tgggatatta cgtcagcgag gacgggtatc ctggctggat gccgcagaaa
     8341 tggacatgga taccccgtga gttacccggc gggcgcgctt ggcgtaatca tggtcatagc
     8401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     8461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     8521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     8581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     8641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     8701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     8761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     8821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     8881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     9001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     9061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     9121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     9181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     9241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     9301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     9361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     9421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     9481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     9541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     9601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     9661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     9721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     9781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     9841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
    10021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
    10081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
    10141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
    10201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
    10261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
    10321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
    10381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
    10441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    10501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
    10561 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
    10621 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
    10681 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
    10741 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
    10801 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
    10861 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
    10921 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
    10981 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
    11041 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
    11101 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
    11161 gcgcgcctcg ttcattcacg tttttgaacc cgtggaggac gggcagactc gcggtgcaaa
    11221 tgtgttttac agcgtgatgg agcagatgaa gatgctcgac acgctgcaga acacgcagct
    11281 agattaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc atgtgtaaaa
    11341 ttgacgcatg tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat
    11401 taagttttat tatatttaca cttacatact aataataaat tcaacaaaca atttatttat
    11461 gtttatttat ttattaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact
    11521 tttatgaggg acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct
    11581 agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc
    11641 gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct
    11701 ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaggc
    11761 ctccacggcc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
    11821 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
    11881 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
    11941 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
    12001 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
    12061 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
    12121 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
    12181 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
    12241 tgtgcagcga tgggggcggg gggggggggg gggg
//
