LOCUS       sequence-429371-       16277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible PE-Plus system for controllable prime editing; this 
            plasmid is used to insert PE-Plus prime editor 
            (PEmax-P2A-hP53DD-IRES-MLH1dn) in one allele of AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16277)
  AUTHORS   Wu Y, Zhong A, Sidharta M, Kim TW, Ramirez B, Persily B, Studer L, 
            Zhou T
  TITLE     Robust and inducible genome editing via an all-in-one prime editor 
            in human pluripotent stem cells.
  JOURNAL   Nat Commun. 2024 Dec 30;15(1):10824. doi: 
            10.1038/s41467-024-55104-1.
  PUBMED    39737975
REFERENCE   2  (bases 1 to 16277)
  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: "Nat 
            Commun."; date: "2024-12-30"; volume: "15(1):10824. doi"; pages: " 
            10.1038/s41467-024-55104-1"
COMMENT     Sequence Label: AAVS1-iPE-Plus
FEATURES             Location/Qualifiers
     source          1..16277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2029..2053)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2215..2296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2252..2271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    2303..2527
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2516..2533
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2565..3590)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2565..3590)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(3600..3653)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3677..3702
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    3709..4512
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(4527..4545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4555..4572)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4555..4571)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4564..4586)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4713..5168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4800..4819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5010..5031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5194..5298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5299..5367,5368..6159)
                     /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     complement(5517..5536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6330..6918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6819..6838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7072..7089
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7206..7227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7242..7247,7248..7265,7266..7272)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7280..7296
                     /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)."
     primer_bind     7285..7307
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7304..7320
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7304..7320
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7339..7359
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7341..7359
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    7378..8214
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        8245..8559
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    8253..8271
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8289..8307
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8324..8342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8360..8378
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8396..8414
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8431..8449
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8467..8485
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     8532..8556
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    8576..8600
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        8635..8653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8703..8723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8724..12824
                     /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
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             12882..12902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             15003..15023
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             15033..15059
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             15069..15125
                     /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"
     misc_feature    join(15450..16024,16025..16027,16028..16036)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(15617..15634)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     15844..15863
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 ccacatctaa gctgcagtct tttgaggatc tggcctctat cagtacctac ggcttcagag
       61 gcgaggccct ggccagcatc agccacgtgg cccatgtgac catcaccacc aaaaccgccg
      121 acggcaaatg cgcttatcgc gctagctaca gcgacggcaa gctgaaagcc ccgccaaagc
      181 cttgcgccgg caaccagggt acacagataa cagtggagga tctgttctac aacatcgcca
      241 cccggagaaa ggccctgaaa aatcccagcg aggagtacgg caagatcctg gaagtcgtcg
      301 gcagatactc cgtgcacaac gccggaatca gctttagcgt aaagaagcag ggagaaaccg
      361 tggccgatgt gcgcaccctg ccaaatgcca gcaccgtgga taacatcaga agcattttcg
      421 gaaatgccgt gtccagagaa ctgatcgaga tcggctgcga agataagacc ctggctttta
      481 agatgaacgg ctacatcagc aacgccaatt actctgtgaa gaagtgcatc tttcttctgt
      541 tcatcaacca cagactggtg gaaagcacca gcctgcggaa agccatcgag acagtgtacg
      601 ccgcctacct gcctaagaac acccacccct tcctgtacct gagcctcgag atcagccctc
      661 agaacgtgga cgtcaatgtg catcctacaa agcacgaggt gcacttcctg cacgaggaaa
      721 gcatcctgga aagagtgcag cagcacattg agagcaagct gctgggctct aacagcagca
      781 gaatgtactt cacacagacc ctgctgcctg gcctggccgg cccctcaggc gaaatggtta
      841 agtccacaac ctctctgacc tcatccagca ccagcggttc ttccgataag gtgtacgccc
      901 accagatggt gcggaccgac tctcgggagc agaagctgga cgcctttctg caacctctga
      961 gcaaacctct gagctctcag cctcaggcca tcgtgaccga ggacaagaca gatatctcct
     1021 ccggccgtgc cagacagcag gacgaagaaa tgctcgagct gccagctcct gccgaggtgg
     1081 ccgccaagaa ccagagcctg gagggagata ccacaaaggg caccagcgaa atgagcgaga
     1141 agcggggccc tacctccagc aaccccagaa aacggcaccg ggaggacagc gacgtggaaa
     1201 tggtggagga cgacagcaga aaggaaatga cagccgcttg tacccctaga agaagaatca
     1261 tcaacctgac ctccgtgctg agcctgcagg aggagatcaa cgagcagggc cacgaggtgc
     1321 tgagagagat gctgcacaat cacagcttcg tgggctgcgt gaaccctcaa tgggccctgg
     1381 ctcagcatca aacaaagctg tacctgctga acaccaccaa gctgagcgaa gagctgttct
     1441 accagatcct catctacgac ttcgccaact tcggcgtgct acgcctgagc gagcccgccc
     1501 ctctgtttga cctggccatg ctggctctgg atagcccaga aagcggctgg acagaagagg
     1561 acggacctaa agaggggctg gctgaataca tcgtggagtt cctgaagaaa aaggccgaga
     1621 tgctggccga ctacttttct ctggaaatcg acgaggaagg caacctgatc ggcctgcctc
     1681 tgctgatcga taactacgtg cctcccctgg aaggcctgcc catcttcatc ctgagactgg
     1741 ctacagaggt gaactgggac gaggaaaagg aatgcttcga gtctctgagc aaggagtgcg
     1801 ccatgttcta tagcatcaga aaacagtaca tctctgaaga gagcactctg tctggccagc
     1861 agagtgaagt gcccggaagc atccccaaca gctggaagtg gaccgtggaa cacatcgtgt
     1921 acaaggccct gcggagccac attctccctc ctaagcactt caccgaggac ggcaacatcc
     1981 tgcagctggc caacctgccc gacctttata aggttttcta acgcgccgac ccagctttct
     2041 tgtacaaagt ggttgatccg gggatcctct agtcagcgtt tatctgcaga attcgccctt
     2101 gtttaaacga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2161 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2281 gcattttttt cactgcctcg acccatagag cccaccgcat ccccagcatg cctgctattg
     2341 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2401 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2461 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2521 ggcacagtcg aggctgatca gcgggtttaa acgggccctc tagactattc ctttgccctc
     2581 ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc
     2641 cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg
     2701 acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag
     2761 ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct
     2821 gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac
     2881 ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc
     2941 cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc
     3001 gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga
     3061 gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg
     3121 ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt
     3181 ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc
     3241 tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca
     3301 ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc
     3361 acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag
     3421 aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg
     3481 aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt
     3541 tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat ctgcagccta
     3601 gggccgggat tctcctccac gtcaccgcat gttagaagac ttcctctgcc ctctccgctg
     3661 ccagatctct cgaggccctg tgggaggaag agaagaggtc agggatctgc cccactgtgg
     3721 ggtggagggg acagataaaa gtacccagaa ccagagccac attaaccggc cctgggaata
     3781 taaggtggtc ccagctcggg gacacaggat ccctggaggc agcaaacatg ctgtcctgaa
     3841 gtggacatag gggcccgggt tggaggaaga agactagctg agctctcgga cccctggaag
     3901 atgccatgac agggggctgg aagagctagc acagactaga gaggtaaggg gggtagggga
     3961 gctgcccaaa tgaaaggagt gagaggtgac ccgaatccac aggagaacgg ggtgtccagg
     4021 caaagaaagc aagaggatgg agaggtggct aaagccaggg agacggggta ctttggggtt
     4081 gtccagaaaa acggtgatga tgcaggccta caagaagggg aggcgggacg caagggagac
     4141 atccgtcgga gaaggccatc ctaagaaacg agagatggca caggccccag aaggagaagg
     4201 aaaagggaac ccagcgagtg aagacggcat ggggttgggt gagggaggag agatgcccgg
     4261 agaggaccca gacacgggga ggatccgctc agaggacatc acgtggtgca gcgccgagaa
     4321 ggaagtgctc cggaaagagc atccttgggc agcaacacag cagagagcaa ggggaagagg
     4381 gagtggagga agacggaacc tgaaggaggc ggcagggaag gatctgggcc agccgtagag
     4441 gtgacccagg ccacaagctg cagacagaaa gcggcacagg cccaggggag agaatgctgg
     4501 tcagagaaag caggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     4561 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4621 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4681 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4741 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4801 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4861 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4921 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4981 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5041 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5101 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5161 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5221 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5281 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5341 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5401 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5461 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5521 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5581 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5641 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5701 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5761 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5821 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5881 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5941 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6001 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6061 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6121 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6181 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6961 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7021 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7081 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7141 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7201 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7261 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7321 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcaga
     7381 gcagagccag gaacccctgt agggaagggg caggagagcc aggggcatga gatggtggac
     7441 gaggaagggg gacagggaag cctgagcgcc tctcctgggc ttgccaagga ctcaaaccca
     7501 gaagcccaga gcagggcctt agggaagcgg gaccctgctc tgggcggagg aatatgtccc
     7561 agatagcact ggggactctt taaggaaaga aggatggaga aagagaaagg gagtagaggc
     7621 ggccacgacc tggtgaacac ctaggacgca ccattctcac aaagggagtt ttccacacgg
     7681 acacccccct cctcaccaca gccctgccag gacggggctg gctactggcc ttatctcaca
     7741 ggtaaaactg acgcacggag gaacaatata aattggggac tagaaaggtg aagagccaaa
     7801 gttagaactc aggaccaact tattctgatt ttgtttttcc aaactgcttc tcctcttggg
     7861 aagtgtaagg aagctgcagc accaggatca gtgaaacgca ccagacagcc gcgtcagagc
     7921 agctcaggtt ctgggagagg gtagcgcagg gtggccactg agaaccgggc aggtcacgca
     7981 tccccccctt ccctcccacc ccctgccaag ctctccctcc caggatcctc tctggctcca
     8041 tcgtaagcaa accttagagg ttctggcaag gagagagatg gctccaggaa atgggggtgt
     8101 gtcaccagat aaggaatctg cctaacagga ggtgggggtt agacccaata tcaggagact
     8161 aggaaggagg aggcctaagg atggggcttt tctgtcacca atcctgtccc tagtttaatt
     8221 aaccctctag agcttagtac tgtcgagttt actccctatc agtgatagag aacgtatgtc
     8281 gagtttactc cctatcagtg atagagaacg atgtcgagtt tactccctat cagtgataga
     8341 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
     8401 atcagtgata gagaacgtat gtcgagttta tccctatcag tgatagagaa cgtatgtcga
     8461 gtttactccc tatcagtgat agagaacgta tgtcgaggta ggcgtgtacg gtgggaggcc
     8521 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagaattcc gatcaacaag
     8581 tttgtacaaa aaagcaggct ccgcggtttt caggttggac cggtgcgcgg ccgctaatac
     8641 gactcactat agggagagcc gccaccatga aacggacagc cgacggaagc gagttcgagt
     8701 caccaaagaa gaagcggaaa gtcgacaaga agtacagcat cggcctggac atcggcacca
     8761 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     8821 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     8881 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     8941 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     9001 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     9061 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     9121 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     9181 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     9241 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     9301 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     9361 ccagactgag caagagcaga aagctggaaa atctgatcgc ccagctgccc ggcgagaaga
     9421 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     9481 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     9541 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     9601 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     9661 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     9721 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     9781 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     9841 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     9901 agagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     9961 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
    10021 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
    10081 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
    10141 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
    10201 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
    10261 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
    10321 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
    10381 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
    10441 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
    10501 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
    10561 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
    10621 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
    10681 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
    10741 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
    10801 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
    10861 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
    10921 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
    10981 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
    11041 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
    11101 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
    11161 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
    11221 ccgactacga tgtggacgct atcgtgcctc agagctttct gaaggacgac tccatcgaca
    11281 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
    11341 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
    11401 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
    11461 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
    11521 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
    11581 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
    11641 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
    11701 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
    11761 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
    11821 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
    11881 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
    11941 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
    12001 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
    12061 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
    12121 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
    12181 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
    12241 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
    12301 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
    12361 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
    12421 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
    12481 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
    12541 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
    12601 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
    12661 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
    12721 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
    12781 gcctgtacga gacacggatc gacctgtctc agctgggagg tgactccggc ggaagctctg
    12841 gtggcagcaa gcggaccgcc gacggctctg aattcgagag ccctaagaag aaaagaaagg
    12901 tgagcggagg ctctagcggc ggaagcaccc tgaacattga agacgagtat agactgcatg
    12961 aaacaagcaa ggaacccgac gtgtccctgg gctccacctg gctgtccgac tttccccagg
    13021 cctgggccga gacaggagga atgggcctgg ccgtgcggca ggcacccctg atcatccctc
    13081 tgaaggccac ctctacaccc gtgagcatca agcagtaccc tatgtctcag gaggccagac
    13141 tgggcatcaa gcctcacatc cagaggctgc tggaccaggg catcctggtg ccatgccaga
    13201 gcccctggaa cacaccactg ctgcccgtga agaagccagg caccaatgac tatagacccg
    13261 tgcaggatct gagagaggtg aacaagaggg tggaggatat ccaccccacc gtgcccaacc
    13321 cttacaatct gctgtccggc ctgccccctt ctcaccagtg gtatacagtg ctggacctga
    13381 aggatgcctt cttttgtctg agactgcacc ctaccagcca gccactgttc gcctttgagt
    13441 ggagggaccc tgagatgggc atctctggcc agctgacctg gacacgcctg cctcagggct
    13501 tcaagaatag cccaacactg tttaacgagg ccctgcaccg cgacctggca gatttccgga
    13561 tccagcaccc agatctgatc ctgctgcagt acgtggacga tctgctgctg gccgccacca
    13621 gcgagctgga ttgccagcag ggaacacgcg ccctgctgca gaccctggga aacctgggat
    13681 atagggcatc cgccaagaag gcccagatct gtcagaagca ggtgaagtac ctgggctatc
    13741 tgctgaagga gggccagaga tggctgacag aggccaggaa ggagacagtg atgggccagc
    13801 caacacccaa gaccccaaga cagctgaggg agttcctggg caaagcagga ttttgcaggc
    13861 tgttcatccc aggattcgca gagatggcag cacctctgta cccactgacc aagccgggca
    13921 ccctgtttaa ttggggccct gaccagcaga aggcctatca ggagatcaag caggccctgc
    13981 tgacagcacc agccctgggc ctgccagacc tgaccaagcc tttcgagctg tttgtggatg
    14041 agaagcaggg ctacgccaag ggcgtgctga cccagaagct gggaccatgg agacggcccg
    14101 tggcctatct gtccaagaag ctggacccag tggcagcagg atggccacca tgcctgagga
    14161 tggtggcagc aatcgccgtg ctgacaaagg atgccggcaa gctgaccatg ggacagccac
    14221 tggtcatcct ggcaccacac gcagtggagg ccctggtgaa gcagcctcca gatcgctggc
    14281 tgtctaacgc ccggatgaca cactaccagg ccctgctgct ggacaccgat cgcgtgcagt
    14341 ttggccctgt ggtggccctg aatccagcca ccctgctgcc tctgccagag gagggcctgc
    14401 agcacaactg tctggacatc ctggcagagg cacacggaac aaggccagac ctgaccgatc
    14461 agcccctgcc tgacgccgat cacacatggt ataccgatgg aagctccctg ctgcaggagg
    14521 gccagaggaa ggcaggagca gcagtgacca cagagacaga agtgatctgg gccaaggccc
    14581 tgccagcagg cacatccgcc cagcgggccg agctgatcgc cctgacccag gccctgaaga
    14641 tggccgaggg caagaagctg aacgtgtaca cagactccag atatgccttc gccaccgcac
    14701 acatccacgg agagatctac aggcgccggg gctggctgac ctctgagggc aaggagatca
    14761 agaacaagga tgagatcctg gccctgctga aggccctgtt tctgcccaag cggctgagca
    14821 tcatccactg tcctggacac cagaagggac actccgccga ggcaaggggc aatcggatgg
    14881 ccgaccaggc cgccagaaag gctgctatta ctgaaactcc cgacacttcc actctgctga
    14941 ttgaaaactc ctccccttct ggcggctcaa aaagaaccgc cgacggcagc gaattcgagt
    15001 ctcccaagaa gaagaggaaa gtcggctctg gccctgccgc taagagagtg aagctggacg
    15061 gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg
    15121 gaccgatgga gcctcaccac gagctgcccc cagggagcac taagcgagca ctgcccaaca
    15181 acaccagctc ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc
    15241 agatccgtgg gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca
    15301 aggatgccca ggctgggaag gagccagggg ggagcagggc tcactccagc cacctgaagt
    15361 ccaaaaaggg tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg
    15421 actcagactg accggtgttt aggcgcgccc ccctctccct cccccccccc taacgttact
    15481 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
    15541 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
    15601 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
    15661 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
    15721 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
    15781 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
    15841 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
    15901 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
    15961 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
    16021 taatatggcc acaaccatga gcttcgttgc tggagtcatc cggagactgg acgagacagt
    16081 ggtgaacaga attgccgccg gcgaggtgat ccagagacct gccaatgcaa taaaggagat
    16141 gatcgagaac tgtctggacg ccaagtccac aagcattcag gtgatcgtga aggagggcgg
    16201 actgaagctg atccagatcc aagacaacgg cacaggcatc agaaaggaag atctggacat
    16261 cgtgtgtgaa cggttca
//
