LOCUS       sequence-346754-       13087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one prime editor piggyBac transposon (xCas9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13087)
  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 13087)
  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 xCas9
FEATURES             Location/Qualifiers
     source          1..13087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        192..440
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     192..212
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     363..382
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        469..544
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        590..969
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1194..2210
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2134..2156
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2218..2237
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2276..2295
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2276..2294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2344..2364
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2365..6465
                     /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
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8638..8658
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8668..8724
                     /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             8725..9321
                     /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(8725..8744)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9221..9241
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             9328..10323
                     /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(10345..10362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10351..10575
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   10759..10821
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(10887..11475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10967..10986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11646..12437,12438..12506))
                     /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     12269..12288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12507..12611)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12679..12697
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12735..12757)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12857..12876
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   12900..12934
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg
       61 ttatttcatg ttctacttac gtgataactt attatatata tattttcttg ttatagatat
      121 catgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggaccc ggtaccaagg
      181 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      241 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
      301 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      361 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      421 tgtggaaagg acgaaacacc gggtcttcgt ggagctcatc gaagacctgt tttagagcta
      481 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      541 gtgcagagac cgaatcgacc agaggtctcg tttttttcta gacgctagcg acattgatta
      601 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      661 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      721 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      781 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      841 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      901 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      961 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     1021 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     1081 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
     1141 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
     1201 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     1261 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     1321 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     1381 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     1441 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     1501 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     1561 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1621 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     1681 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     1741 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     1801 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     1861 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1921 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     1981 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     2041 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     2101 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     2161 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     2221 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgatagcg gccgctaata
     2281 cgactcacta tagggagagc cgccaccatg aaacggacag ccgacggaag cgagttcgag
     2341 tcaccaaaga agaagcggaa agtcgacaag aagtacagca tcggcctgga catcggcacc
     2401 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     2461 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     2521 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     2581 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     2641 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     2701 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     2761 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     2821 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     2881 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     2941 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     3001 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     3061 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     3121 agcaacttcg acctggccga ggataccaaa ctgcagctga gcaaggacac ctacgacgac
     3181 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     3241 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     3301 aaggcccccc tgagcgcctc tatgatcaag ctgtacgacg agcaccacca ggacctgacc
     3361 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     3421 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     3481 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     3541 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcatcat cccccaccag
     3601 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     3661 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     3721 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     3781 accccctgga acttcgagaa ggtggtggac aagggcgctt ccgcccagag cttcatcgag
     3841 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     3901 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     3961 atgagaaagc ccgccttcct gagcggcgac cagaaaaagg ccatcgtgga cctgctgttc
     4021 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     4081 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     4141 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     4201 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     4261 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     4321 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     4381 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     4441 atccagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     4501 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     4561 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     4621 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     4681 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     4741 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     4801 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     4861 tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     4921 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     4981 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     5041 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     5101 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     5161 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     5221 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     5281 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     5341 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     5401 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     5461 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     5521 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     5581 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     5641 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     5701 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     5761 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     5821 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     5881 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     5941 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     6001 ctggcctctg ccggcgtgct gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     6061 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     6121 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     6181 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     6241 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     6301 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     6361 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     6421 ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg aggatctagc
     6481 ggaggatcct ctggcagcga gacaccagga acaagcgagt cagcaacacc agagagcagt
     6541 ggcggcagca gcggcggcag cagcacccta aatatagaag atgagtatcg gctacatgaa
     6601 acctcaaaag agccagatgt ttctctaggg tccacatggc tgtctgattt tcctcaggcc
     6661 tgggcggaaa ccgggggcat gggactggca gttcgccaag ctcctctgat catacctctg
     6721 aaagcaacct ctacccccgt gtccataaaa caatacccca tgtcacaaga agccagactg
     6781 gggatcaagc cccacataca gagactgttg gaccagggaa tactggtacc ctgccagtcc
     6841 ccctggaaca cgcccctgct acccgttaag aaaccaggga ctaatgatta taggcctgtc
     6901 caggatctga gagaagtcaa caagcgggtg gaggacatcc accccaccgt gcccaaccct
     6961 tacaacctct tgagcgggct cccaccgtcc caccagtggt acactgtgct tgatttaaag
     7021 gatgcctttt tctgcctgag actccacccc accagtcagc ctctcttcgc ctttgagtgg
     7081 agagatccag agatgggaat ctcaggacaa ttgacctgga ccagactccc acagggtttc
     7141 aaaaacagtc ccaccctgtt taatgaggca ctgcacaggg acctagcaga cttcagaatc
     7201 cagcacccag acttgatcct gctacagtac gtggatgact tactgctggc cgccacttct
     7261 gagctagact gccaacaagg tactcgggcc ctgttacaaa ccctagggaa cctcgggtat
     7321 cgggcctcgg ccaagaaagc ccaaatttgc cagaaacagg tcaagtatct ggggtatctt
     7381 ctaaaagagg gtcagagatg gctgactgag gccagaaaag agactgtgat ggggcagcct
     7441 actcccaaga cccctcgaca actaagggag ttcctaggga aggcaggctt ctgtcgcctc
     7501 ttcatccctg ggtttgcaga aatggcagcc cccctgtacc ctctcaccaa accggggact
     7561 ctgtttaatt ggggcccaga ccaacaaaag gcctatcaag aaatcaagca agctcttcta
     7621 actgccccag ccctggggtt gccagatttg actaagccct ttgaactctt tgtcgacgag
     7681 aagcagggct acgccaaagg tgtcctaacg caaaaactgg gaccttggcg tcggccggtg
     7741 gcctacctgt ccaaaaagct agacccagta gcagctgggt ggcccccttg cctacggatg
     7801 gtagcagcca ttgccgtact gacaaaggat gcaggcaagc taaccatggg acagccacta
     7861 gtcattctgg ccccccatgc agtagaggca ctagtcaaac aaccccccga ccgctggctt
     7921 tccaacgccc ggatgactca ctatcaggcc ttgcttttgg acacggaccg ggtccagttc
     7981 ggcccggtgg tagccctgaa cccggctacg ctgctcccac tgcctgagga agggctgcaa
     8041 cacaactgcc ttgatatcct ggccgaagcc cacggaaccc gacccgacct aacggaccag
     8101 ccgctcccag acgccgacca cacctggtac acggatggaa gcagtctctt acaagaggga
     8161 cagcgtaagg cgggagctgc ggtgaccacc gagacagagg taatctgggc taaagccctg
     8221 ccagccggga catccgctca gcgggctgaa ctgatagcac tcacccaggc cctaaagatg
     8281 gcagaaggta agaagctaaa tgtttatact gatagccgtt atgcttttgc tactgcccat
     8341 atccatggag aaatatacag aaggcgtggg tggctcacat cagaaggcaa agagatcaaa
     8401 aataaagacg agatcttggc cctactaaaa gccctctttc tgcccaaaag acttagcata
     8461 atccattgtc caggacatca aaagggacac agcgccgagg ctagaggcaa ccggatggct
     8521 gaccaagcgg cccgaaaggc agccatcaca gagactccag acacctctac cctcctcata
     8581 gaaaattcat caccctctgg cggctcaaaa agaaccgccg acggcagcga attcgagccc
     8641 aagaagaaga ggaaagtcgg ctccggcgct accaatttct ccctgctcaa gcaagccgga
     8701 gacgtcgagg aaaacccagg cccaatgacc gagtacaagc ccacggtgcg cctcgccacc
     8761 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     8821 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     8881 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     8941 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     9001 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     9061 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     9121 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     9181 cgcgccgggg tgcccgcctt cctggagaca tccgcgcccc gcaacctccc cttctacgag
     9241 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     9301 atgacccgca agcccggtgc cggctccatg cccacgctac tgcgggttta tatagacggt
     9361 cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac
     9421 gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc ttccgagaca
     9481 atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac
     9541 gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac
     9601 gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc
     9661 ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga
     9721 taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg
     9781 accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca catcgaccgc
     9841 ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc
     9901 cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg gtcgtggcgg
     9961 gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc
    10021 aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag
    10081 ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga cgtcttggcc
    10141 aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc
    10201 cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc
    10261 tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac
    10321 tgagctctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
    10381 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
    10441 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
    10501 gggtggggca ggacagcaag ggggaggatt gggaggacaa tagcaggcat gctggggatg
    10561 cggtgggctc tatggggcgc gcctaaaagt tttgttactt tatagaagaa attttgagtt
    10621 tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat ttattattag
    10681 tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat aaacctcgat
    10741 atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac
    10801 aatatgatta tctttctagg gttaaacatg tgagcaaaag gccagcaaaa ggccaggaac
    10861 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    10921 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    10981 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    11041 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    11101 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    11161 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    11221 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    11281 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    11341 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    11401 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    11461 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    11521 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    11581 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    11641 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    11701 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    11761 ggccccagtg ctgcaatgat accgcgactc ccacgctcac cggctccaga tttatcagca
    11821 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    11881 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    11941 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    12001 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    12061 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    12121 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    12181 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    12241 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    12301 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    12361 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    12421 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    12481 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    12541 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    12601 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
    12661 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
    12721 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
    12781 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
    12841 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca atttgttaac
    12901 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
    12961 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
    13021 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
    13081 agtcaaa
//
