LOCUS       sequence-320034-       13140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one prime editor piggyBac transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13140)
  AUTHORS   Eggenschwiler R, Gschwendtberger T, Felski C, Jahn C, Langer F, 
            Sterneckert J, Hermann A, Luhmann J, Steinemann D, Haase A, Martin 
            U, Petri S, Cantz T
  TITLE     A selectable all-in-one CRISPR prime editing piggyBac transposon 
            allows for highly efficient gene editing in human cell lines.
  JOURNAL   Sci Rep. 2021 Nov 12;11(1):22154. doi: 10.1038/s41598-021-01689-2.
  PUBMED    34773059
REFERENCE   2  (bases 1 to 13140)
  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: "Sci Rep."; 
            date: "2021-11-12"; volume: "11(1):22154. doi"; pages: " 
            10.1038/s41598-021-01689-2"
COMMENT     Sequence Label: PB-PE
FEATURES             Location/Qualifiers
     source          1..13140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..265
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     17..37
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     188..207
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        294..369
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        415..794
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        796..1071
                     /label=chicken beta-actin promoter
     intron          1072..2088
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2012..2034
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2096..2115
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2154..2173
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2154..2172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2222..2242
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2243..4756,4757..4759,4760..6343)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2243..6343
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8516..8536
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8546..8602
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8603..9199
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8603..8622)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9099..9119
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             9206..10201
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(10223..10240)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10229..10453
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   10637..10699
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(10765..11353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10845..10864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11524..12315,12316..12384))
                     /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     12147..12166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12385..12489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12557..12575
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12613..12635)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12735..12754
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   12778..12812
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata
       61 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
      121 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      181 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      241 tatcttgtgg aaaggacgaa acaccgggtc ttcgtggagc tcatcgaaga cctgttttag
      301 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      361 agtcggtgca gagaccgaat cgaccagagg tctcgttttt ttctagacgc tagcgacatt
      421 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      481 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      541 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      601 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      661 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      721 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      781 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
      841 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
      901 cggggggggg gggggggcgc gccrgggggg gggggggggg gggggggggg gggsggggsg
      961 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     1021 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     1081 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     1141 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     1201 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     1261 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     1321 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     1381 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     1441 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     1501 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     1561 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     1621 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     1681 cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     1741 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     1801 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     1861 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1921 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     1981 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     2041 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     2101 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgatagcggc cgctaatacg
     2161 actcactata gggagagccg ccaccatgaa acggacagcc gacggaagcg agttcgagtc
     2221 accaaagaag aagcggaaag tcgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     2281 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     2341 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     2401 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     2461 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     2521 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     2581 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     2641 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     2701 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     2761 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     2821 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     2881 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     2941 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     3001 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     3061 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     3121 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     3181 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     3241 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     3301 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     3361 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     3421 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     3481 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     3541 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     3601 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     3661 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     3721 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     3781 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     3841 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     3901 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     3961 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     4021 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     4081 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     4141 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     4201 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     4261 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     4321 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     4381 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     4441 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     4501 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     4561 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     4621 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     4681 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     4741 cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     4801 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     4861 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     4921 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     4981 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     5041 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     5101 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     5161 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     5221 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     5281 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     5341 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     5401 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     5461 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     5521 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     5581 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     5641 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     5701 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     5761 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     5821 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     5881 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     5941 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     6001 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     6061 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     6121 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     6181 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     6241 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     6301 cctgtacgag acacggatcg acctgtctca gctgggaggt gactctggag gatctagcgg
     6361 aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag agagcagtgg
     6421 cggcagcagc ggcggcagca gcaccctaaa tatagaagat gagtatcggc tacatgaaac
     6481 ctcaaaagag ccagatgttt ctctagggtc cacatggctg tctgattttc ctcaggcctg
     6541 ggcggaaacc gggggcatgg gactggcagt tcgccaagct cctctgatca tacctctgaa
     6601 agcaacctct acccccgtgt ccataaaaca ataccccatg tcacaagaag ccagactggg
     6661 gatcaagccc cacatacaga gactgttgga ccagggaata ctggtaccct gccagtcccc
     6721 ctggaacacg cccctgctac ccgttaagaa accagggact aatgattata ggcctgtcca
     6781 ggatctgaga gaagtcaaca agcgggtgga ggacatccac cccaccgtgc ccaaccctta
     6841 caacctcttg agcgggctcc caccgtccca ccagtggtac actgtgcttg atttaaagga
     6901 tgcctttttc tgcctgagac tccaccccac cagtcagcct ctcttcgcct ttgagtggag
     6961 agatccagag atgggaatct caggacaatt gacctggacc agactcccac agggtttcaa
     7021 aaacagtccc accctgttta atgaggcact gcacagggac ctagcagact tcagaatcca
     7081 gcacccagac ttgatcctgc tacagtacgt ggatgactta ctgctggccg ccacttctga
     7141 gctagactgc caacaaggta ctcgggccct gttacaaacc ctagggaacc tcgggtatcg
     7201 ggcctcggcc aagaaagccc aaatttgcca gaaacaggtc aagtatctgg ggtatcttct
     7261 aaaagagggt cagagatggc tgactgaggc cagaaaagag actgtgatgg ggcagcctac
     7321 tcccaagacc cctcgacaac taagggagtt cctagggaag gcaggcttct gtcgcctctt
     7381 catccctggg tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac cggggactct
     7441 gtttaattgg ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag ctcttctaac
     7501 tgccccagcc ctggggttgc cagatttgac taagcccttt gaactctttg tcgacgagaa
     7561 gcagggctac gccaaaggtg tcctaacgca aaaactggga ccttggcgtc ggccggtggc
     7621 ctacctgtcc aaaaagctag acccagtagc agctgggtgg cccccttgcc tacggatggt
     7681 agcagccatt gccgtactga caaaggatgc aggcaagcta accatgggac agccactagt
     7741 cattctggcc ccccatgcag tagaggcact agtcaaacaa ccccccgacc gctggctttc
     7801 caacgcccgg atgactcact atcaggcctt gcttttggac acggaccggg tccagttcgg
     7861 cccggtggta gccctgaacc cggctacgct gctcccactg cctgaggaag ggctgcaaca
     7921 caactgcctt gatatcctgg ccgaagccca cggaacccga cccgacctaa cggaccagcc
     7981 gctcccagac gccgaccaca cctggtacac ggatggaagc agtctcttac aagagggaca
     8041 gcgtaaggcg ggagctgcgg tgaccaccga gacagaggta atctgggcta aagccctgcc
     8101 agccgggaca tccgctcagc gggctgaact gatagcactc acccaggccc taaagatggc
     8161 agaaggtaag aagctaaatg tttatactga tagccgttat gcttttgcta ctgcccatat
     8221 ccatggagaa atatacagaa ggcgtgggtg gctcacatca gaaggcaaag agatcaaaaa
     8281 taaagacgag atcttggccc tactaaaagc cctctttctg cccaaaagac ttagcataat
     8341 ccattgtcca ggacatcaaa agggacacag cgccgaggct agaggcaacc ggatggctga
     8401 ccaagcggcc cgaaaggcag ccatcacaga gactccagac acctctaccc tcctcataga
     8461 aaattcatca ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa
     8521 gaagaagagg aaagtcggct ccggcgctac caatttctcc ctgctcaagc aagccggaga
     8581 cgtcgaggaa aacccaggcc caatgaccga gtacaagccc acggtgcgcc tcgccacccg
     8641 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     8701 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     8761 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     8821 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     8881 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     8941 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     9001 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     9061 cgccggggtg cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg
     9121 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     9181 gacccgcaag cccggtgccg gctccatgcc cacgctactg cgggtttata tagacggtcc
     9241 tcacgggatg gggaaaacca ccaccacgca actgctggtg gccctgggtt cgcgcgacga
     9301 tatcgtctac gtacccgagc cgatgactta ctggcaggtg ctgggggctt ccgagacaat
     9361 cgcgaacatc tacaccacac aacaccgcct cgaccagggt gagatatcgg ccggggacgc
     9421 ggcggtggta atgacaagcg cccagataac aatgggcatg ccttatgccg tgaccgacgc
     9481 cgttctggct cctcatatcg ggggggaggc tgggagctca catgccccgc ccccggccct
     9541 caccctcatc ttcgaccgcc atcccatcgc cgccctcctg tgctacccgg ccgcgcgata
     9601 ccttatgggc agcatgaccc cccaggccgt gctggcgttc gtggccctca tcccgccgac
     9661 cttgcccggc acaaacatcg tgttgggggc ccttccggag gacagacaca tcgaccgcct
     9721 ggccaaacgc cagcgccccg gcgagcggct tgacctggct atgctggccg cgattcgccg
     9781 cgtttacggg ctgcttgcca atacggtgcg gtatctgcag ggcggcgggt cgtggcggga
     9841 ggattgggga cagctttcgg ggacggccgt gccgccccag ggtgccgagc cccagagcaa
     9901 cgcgggccca cgaccccata tcggggacac gttatttacc ctgtttcggg cccccgagtt
     9961 gctggccccc aacggcgacc tgtacaacgt gtttgcctgg gccttggacg tcttggccaa
    10021 acgcctccgt cccatgcacg tctttatcct ggattacgac caatcgcccg ccggctgccg
    10081 ggacgccctg ctgcaactta cctccgggat ggtccagacc cacgtcacca cccccggctc
    10141 cataccgacg atctgcgacc tggcgcgcac gtttgcccgg gagatggggg aggctaactg
    10201 agctctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
    10261 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
    10321 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
    10381 gtggggcagg acagcaaggg ggaggattgg gaggacaata gcaggcatgc tggggatgcg
    10441 gtgggctcta tggggcgcgc ctaaaagttt tgttacttta tagaagaaat tttgagtttt
    10501 tgtttttttt taataaataa ataaacataa ataaattgtt tgttgaattt attattagta
    10561 tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa acctcgatat
    10621 acagaccgat aaaacacatg cgtcaatttt acgcatgatt atctttaacg tacgtcacaa
    10681 tatgattatc tttctagggt taaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    10741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    10801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    10861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    10921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    10981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    11041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    11101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    11161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
    11221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    11281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    11341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    11401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
    11461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
    11521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
    11581 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
    11641 ccccagtgct gcaatgatac cgcgactccc acgctcaccg gctccagatt tatcagcaat
    11701 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    11761 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    11821 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    11881 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    11941 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    12001 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    12061 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    12121 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    12181 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    12241 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    12301 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    12361 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    12421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    12481 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
    12541 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
    12601 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
    12661 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
    12721 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccaat ttgttaaccc
    12781 tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct
    12841 aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg
    12901 tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag
    12961 tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta
    13021 tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata
    13081 gatatcatgt acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg gacccggtac
//
