LOCUS       sequence-354084-       12239 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 12239)
  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 12239)
  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_EYFP
FEATURES             Location/Qualifiers
     source          1..12239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          117..1133
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1057..1079
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1141..1160
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1230..1239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1236..1262,1263..1265,1266..3749,3750..3755,3756..3821,
                     3822..3824,3825..5339)
                     /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             5352..5372
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5433..5627
                     /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(5682..5701)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5726..5774
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6006..6217
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6227..6236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6233..6235,6236..6238,6239..6952)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6278..6299)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6539..6558)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6886..6907
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6956..7077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6993..7012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7047..7066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7840..7874
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8365..8381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8365..8381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8378..8400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8389..8405
                     /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(8413..8419,8420..8437,8438..8443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8458..8479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8596..8613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8767..9355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8847..8866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9526..10317,10318..10386))
                     /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     10149..10168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10387..10491)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10517..10972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10654..10675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10866..10885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11099..11121
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11113..11130
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11114..11130
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11260..11322
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11743..12122
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      301 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
      361 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
      421 ggcgcggcgc ggggcttttt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
      481 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
      541 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
      601 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
      661 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
      721 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
      781 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
      841 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
      901 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
      961 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1021 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1081 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1141 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaatctaga taccctcgta
     1201 aaggtctaga gctagcggat cgaacccttg ccaccatgga caagaagtac tccattgggc
     1261 tcgctatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac aaggtgccga
     1321 gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg
     1381 gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac
     1441 ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg
     1501 agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg
     1561 aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc
     1621 atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg
     1681 ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc
     1741 tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg
     1801 ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca
     1861 aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc
     1921 tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga
     1981 cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag
     2041 acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc
     2101 tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga
     2161 acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc
     2221 accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg
     2281 aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc
     2341 aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc
     2401 tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa
     2461 gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt
     2521 tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac
     2581 cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat
     2641 cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc
     2701 agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc
     2761 ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat
     2821 acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg
     2881 tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt
     2941 tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg
     3001 catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca
     3061 atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata
     3121 gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga
     3181 aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg
     3241 ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg
     3301 ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc
     3361 agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag
     3421 gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca
     3481 aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa
     3541 ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa
     3601 aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg
     3661 agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg
     3721 acatcaatcg gctctccgac tacgacgtgg ctgctatcgt gccccagtct tttctcaaag
     3781 atgattctat tgataataaa gtgttgacaa gatccgataa agctagaggg aagagtgata
     3841 acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg
     3901 ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt
     3961 ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca
     4021 agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac
     4081 tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg
     4141 actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc
     4201 tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg
     4261 tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa
     4321 taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg
     4381 agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa
     4441 caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca
     4501 tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa
     4561 gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca
     4621 agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag
     4681 tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca
     4741 tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag
     4801 aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg
     4861 gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc
     4921 cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc
     4981 ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga
     5041 tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata
     5101 aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca
     5161 ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca
     5221 ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc
     5281 atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagaca
     5341 gcagggctga ccccaagaag aagaggaagg tgagtggtgg aggaagtggc gggtcagggt
     5401 cgatggacgc gaaatcactt acggcatggt cgagaacact ggttacgttc aaggacgtgt
     5461 ttgtggactt tacacgtgag gagtggaaat tgctggatac tgcgcaacaa attgtgtatc
     5521 gaaatgtcat gcttgagaat tacaagaacc tcgtcagtct cggataccag ttgacgaaac
     5581 cggatgtgat ccttaggctc gaaaaggggg aagaaccttg gctggtatag aagaattcga
     5641 tccctaccgg ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca
     5701 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     5761 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5821 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5881 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5941 agatctgcga aatcggatcc gcggccgcac tgattctagc tgcagaggct ccggtgcccg
     6001 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     6061 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     6121 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     6181 cgttcttttt cgcaacgggt ttgccgccag aacacaggat cctcgagcca ccatggtgag
     6241 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     6301 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     6361 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     6421 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     6481 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     6541 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     6601 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     6661 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     6721 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     6781 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     6841 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     6901 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagttaactt
     6961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7021 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7081 tgtctggaat tgactcaaat gatgtcaatt agtctatcag aagctcatct ggtctccctt
     7141 ccgggggaca agacatccct gtttaatatt taaacagcag tgttcccaaa ctgggttctt
     7201 atatcccttg ctctggtcaa ccaggttgca gggtttcctg tcctcacagg aacgaagtcc
     7261 ctaaagaaac agtggcagcc aggtttagcc ccggaattga ctggattcct tttttagggc
     7321 ccattggtat ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga
     7381 gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac
     7441 gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc
     7501 tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg
     7561 tccctgatat ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg
     7621 aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg
     7681 cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca
     7741 agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat
     7801 tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga
     7861 ctatctttct agggttaatc tagctgcatc aggatcatat cgtcgggtct tttttccggc
     7921 tcagtcatcg cccaagctgg cgctatctgg gcatcgggga ggaagaagcc cgtgcctttt
     7981 cccgcgaggt tgaagcggca tggaaagagt ttgccgagga tgactgctgc tgcattgacg
     8041 ttgagcgaaa acgcacgttt accatgatga ttcgggaagg tgtggccatg cacgccttta
     8101 acggtgaact gttcgttcag gccacctggg ataccagttc gtcgcggctt ttccggacac
     8161 agttccggat ggtcagcccg aagcgcatca gcaacccgaa caataccggc gacagccgga
     8221 actgccgtgc cggtgtgcag attaatgaca gcggtgcggc gctgggatat tacgtcagcg
     8281 aggacgggta tcctggctgg atgccgcaga aatggacatg gataccccgt gagttacccg
     8341 gcgggcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     8401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     8461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     8521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     8581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     8641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     8701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     8761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     8821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     8881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     8941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     9001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     9061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     9121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     9181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     9241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     9301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     9361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     9421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     9481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     9541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     9601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     9661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     9721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     9781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     9841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    10021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    10081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    10141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    10201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    10261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    10321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    10381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    10441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    10501 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
    10561 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
    10621 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
    10681 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
    10741 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
    10801 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
    10861 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
    10921 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
    10981 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
    11041 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
    11101 cagtcacgac gttgtaaaac gacggccagt gagcgcgcct cgttcattca cgtttttgaa
    11161 cccgtggagg acgggcagac tcgcggtgca aatgtgtttt acagcgtgat ggagcagatg
    11221 aagatgctcg acacgctgca gaacacgcag ctagattaac cctagaaaga taatcatatt
    11281 gtgacgtacg ttaaagataa tcatgtgtaa aattgacgca tgtgttttat cggtctgtat
    11341 atcgaggttt atttattaat ttgaatagat attaagtttt attatattta cacttacata
    11401 ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa aaaaacaaaa
    11461 actcaaaatt tcttctataa agtaacaaaa cttttatgag ggacagcccc cccccaaagc
    11521 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
    11581 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
    11641 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
    11701 tgcagacacc tggggggata cggggaaaag gcctccacgg ccgacattga ttattgacta
    11761 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
    11821 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
    11881 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
    11941 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
    12001 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
    12061 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
    12121 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
    12181 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg ggggggggg
//
