LOCUS       sequence-354088-       12303 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 12303)
  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 12303)
  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_KanR neo
FEATURES             Location/Qualifiers
     source          1..12303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          110..1126
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1050..1072
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1134..1153
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1223..1232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1229..1255,1256..1258,1259..3742,3743..3748,3749..3814,
                     3815..3817,3818..5332)
                     /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             5345..5365
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5426..5620
                     /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(5675..5694)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5719..5767
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5999..6210
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6226..7020
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6226..7020
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6280..6299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6890..6909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7024..7145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7061..7080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7115..7134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7908..7942
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8433..8449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8433..8449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8446..8468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8457..8473
                     /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(8481..8487,8488..8505,8506..8511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8526..8547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8664..8681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8835..9423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8915..8934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9594..10385,10386..10454))
                     /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     10217..10236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10455..10559)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10585..11040)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10722..10743)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10934..10953
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11167..11189
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11181..11198
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11182..11198
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11328..11390
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11811..12190
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      241 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      301 gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg
      361 cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
      421 cgcggggctt tttgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc
      481 gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
      541 gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg
      601 agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct
      661 cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg
      721 ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
      781 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
      841 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
      901 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
      961 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1021 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1081 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1141 gctggttatt gtgctgtctc atcattttgg caaagaatct agataccctc gtaaaggtct
     1201 agagctagcg gatcgaaccc ttgccaccat ggacaagaag tactccattg ggctcgctat
     1261 cggcacaaac agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa
     1321 attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggcgccct
     1381 cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag
     1441 atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc
     1501 taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa
     1561 aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa
     1621 gtacccaacc atatatcatc tgaggaagaa gcttgtagac agtactgata aggctgactt
     1681 gcggttgatc tatctcgcgc tggcgcatat gatcaaattt cggggacact tcctcatcga
     1741 gggggacctg aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac
     1801 ttacaatcag cttttcgaag agaacccgat caacgcatcc ggagttgacg ccaaagcaat
     1861 cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg
     1921 ggagaagaag aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa
     1981 ctttaaatct aacttcgacc tggccgaaga tgccaagctt caactgagca aagacaccta
     2041 cgatgatgat ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt
     2101 ggcggcaaag aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga
     2161 gatcaccaaa gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga
     2221 cttgactttg ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt
     2281 cttcgatcag tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga
     2341 attttacaaa tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt
     2401 aaagcttaac agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc
     2461 ccaccagatt cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc
     2521 ctttttgaaa gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta
     2581 tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg atgactcgca aatcagaaga
     2641 gaccatcact ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt
     2701 catcgaaagg atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca
     2761 ctctctgctg tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac
     2821 agaagggatg agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct
     2881 cctcttcaag acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa
     2941 gattgaatgt ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct
     3001 gggaacgtat cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga
     3061 gaacgaggac attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat
     3121 gattgaagaa cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct
     3181 caagaggcgc cgatatacag gatgggggcg gctgtcaaga aaactgatca atgggatccg
     3241 agacaagcag agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaaccg
     3301 gaacttcatg cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc
     3361 acaagtttct ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc
     3421 agctatcaaa aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat
     3481 gggaaggcat aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca
     3541 gaagggacag aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact
     3601 ggggtcccaa atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct
     3661 ctacctgtac tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa
     3721 tcggctctcc gactacgacg tggctgctat cgtgccccag tcttttctca aagatgattc
     3781 tattgataat aaagtgttga caagatccga taaagctaga gggaagagtg ataacgtccc
     3841 ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact
     3901 gatcacacaa cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt
     3961 ggataaagcc ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt
     4021 ggcccaaatt ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg
     4081 agaggtgaaa gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca
     4141 gttttataag gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc
     4201 agtggtaggc actgcactta tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg
     4261 agactataaa gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa
     4321 ggccaccgct aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac
     4381 actggccaat ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga
     4441 aatcgtgtgg gacaagggta gggatttcgc gacagtccgg aaggtcctgt ccatgccgca
     4501 ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct
     4561 cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata
     4621 cggcggattc gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa
     4681 agggaagtct aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg
     4741 atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata aagaggtcaa
     4801 aaaagacctc atcattaagc ttcccaagta ctctctcttt gagcttgaaa acggccggaa
     4861 acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa
     4921 atacgttaat ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga
     4981 taatgagcag aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga
     5041 gcaaataagc gaattctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct
     5101 ttctgcttac aataagcaca gggataagcc catcagggag caggcagaaa acattatcca
     5161 cttgtttact ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat
     5221 agacagaaag cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc
     5281 aattacgggg ctctatgaaa caagaatcga cctctctcag ctcggtggag acagcagggc
     5341 tgaccccaag aagaagagga aggtgagtgg tggaggaagt ggcgggtcag ggtcgatgga
     5401 cgcgaaatca cttacggcat ggtcgagaac actggttacg ttcaaggacg tgtttgtgga
     5461 ctttacacgt gaggagtgga aattgctgga tactgcgcaa caaattgtgt atcgaaatgt
     5521 catgcttgag aattacaaga acctcgtcag tctcggatac cagttgacga aaccggatgt
     5581 gatccttagg ctcgaaaagg gggaagaacc ttggctggta tagaagaatt cgatccctac
     5641 cggttagtaa tgagtttaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     5701 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     5761 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5821 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5881 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     5941 cgaaatcgga tccgcggccg cactgattct agctgcagag gctccggtgc ccgtcagtgg
     6001 gcagagcgca catcgcccac agcccccgag aagttggggg gaggggtcgg caattgaacc
     6061 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     6121 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     6181 tttcgcaacg ggtttgccgc cagaacacag gatcctcgag ccaccatgat tgaacaagat
     6241 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6301 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6361 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6421 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6481 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6541 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6601 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6661 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6721 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6781 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6841 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6901 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6961 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7021 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7081 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7141 tcttatcatg tctggaattg actcaaatga tgtcaattag tctatcagaa gctcatctgg
     7201 tctcccttcc gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact
     7261 gggttcttat atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa
     7321 cgaagtccct aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt
     7381 tttagggccc attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
     7441 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
     7501 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
     7561 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
     7621 gggggctgtc cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt
     7681 agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga
     7741 taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg
     7801 cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag
     7861 cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt
     7921 tacgcagact atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt
     7981 tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg
     8041 tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg
     8101 cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca
     8161 cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt
     8221 ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga
     8281 cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta
     8341 cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga
     8401 gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     8461 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     8521 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     8581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     8641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     8701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     8761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     8821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     8881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     8941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     9001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     9061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     9121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     9181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     9241 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     9301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     9361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     9421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     9481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     9541 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     9601 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     9661 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     9721 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     9781 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     9841 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     9901 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     9961 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    10021 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    10081 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    10141 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    10201 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    10261 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    10321 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    10381 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    10441 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    10501 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    10561 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
    10621 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
    10681 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
    10741 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
    10801 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
    10861 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
    10921 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
    10981 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
    11041 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
    11101 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
    11161 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcctcg ttcattcacg
    11221 tttttgaacc cgtggaggac gggcagactc gcggtgcaaa tgtgttttac agcgtgatgg
    11281 agcagatgaa gatgctcgac acgctgcaga acacgcagct agattaaccc tagaaagata
    11341 atcatattgt gacgtacgtt aaagataatc atgtgtaaaa ttgacgcatg tgttttatcg
    11401 gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca
    11461 cttacatact aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa
    11521 aaacaaaaac tcaaaatttc ttctataaag taacaaaact tttatgaggg acagcccccc
    11581 cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc
    11641 ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga
    11701 cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc
    11761 tttgagcctg cagacacctg gggggatacg gggaaaaggc ctccacggcc gacattgatt
    11821 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
    11881 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
    11941 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
    12001 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
    12061 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
    12121 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
    12181 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
    12241 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
    12301 ggg
//
