LOCUS       sequence-354057-       12263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12263)
  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 12263)
  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_Hygro
FEATURES             Location/Qualifiers
     source          1..12263
                     /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"
     primer_bind     complement(5390..5409)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5434..5482
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5714..5925
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5935..5944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5956..6975
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             5956..6975
                     /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
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     polyA_signal    6979..7100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7016..7035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7070..7089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7863..7897
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8388..8404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8388..8404)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8401..8423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8412..8428
                     /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(8436..8442,8443..8460,8461..8466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8481..8502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8619..8636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8790..9378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8870..8889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9549..10340,10341..10409))
                     /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     10172..10191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10410..10514)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10540..10995)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10677..10698)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10889..10908
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11122..11144
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11136..11153
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11137..11153
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11283..11345
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11766..12145
                     /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 aggtgtgagt ttaaacgggg gaggctaact gaaacacgga
     5401 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5461 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5521 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5581 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5641 gccatagcag atctgcgaaa tcggatccgc ggccgcactg attctagctg cagaggctcc
     5701 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     5761 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     5821 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     5881 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggatcc tcgagccacc
     5941 atggatagat ccggaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     6001 aagttcgaca gcgtttccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     6061 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     6121 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     6181 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     6241 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     6301 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     6361 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     6421 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     6481 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     6541 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     6601 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     6661 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     6721 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     6781 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     6841 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     6901 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     6961 agggcaaagg aataggttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     7021 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     7081 aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca attagtctat
     7141 cagaagctca tctggtctcc cttccggggg acaagacatc cctgtttaat atttaaacag
     7201 cagtgttccc aaactgggtt cttatatccc ttgctctggt caaccaggtt gcagggtttc
     7261 ctgtcctcac aggaacgaag tccctaaaga aacagtggca gccaggttta gccccggaat
     7321 tgactggatt ccttttttag ggcccattgg tatggctttt tccccgtatc cccccaggtg
     7381 tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc
     7441 ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag cgccggaccg
     7501 gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt aattacatcc
     7561 ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat atatataata
     7621 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     7681 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     7741 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     7801 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     7861 tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc atcaggatca
     7921 tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc tgggcatcgg
     7981 ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag agtttgccga
     8041 ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga tgattcggga
     8101 aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct gggataccag
     8161 ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca tcagcaaccc
     8221 gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg acagcggtgc
     8281 ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc agaaatggac
     8341 atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc atagctgttt
     8401 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8461 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8521 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8581 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     9001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    10021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    10081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10501 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
    10561 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
    10621 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
    10681 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
    10741 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
    10801 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
    10861 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
    10921 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
    10981 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
    11041 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
    11101 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
    11161 cctcgttcat tcacgttttt gaacccgtgg aggacgggca gactcgcggt gcaaatgtgt
    11221 tttacagcgt gatggagcag atgaagatgc tcgacacgct gcagaacacg cagctagatt
    11281 aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgtg taaaattgac
    11341 gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata gatattaagt
    11401 tttattatat ttacacttac atactaataa taaattcaac aaacaattta tttatgttta
    11461 tttatttatt aaaaaaaaca aaaactcaaa atttcttcta taaagtaaca aaacttttat
    11521 gagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
    11581 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
    11641 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
    11701 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaggcctcca
    11761 cggccgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
    11821 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
    11881 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
    11941 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
    12001 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
    12061 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
    12121 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
    12181 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
    12241 agcgatgggg gcgggggggg ggg
//
