LOCUS       sequence-429523-       13049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible PEmax system for controllable prime editing; this plasmid 
            is used to insert PEmax prime editor in one allele of AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13049)
  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 13049)
  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-iPEmax-Hygro
FEATURES             Location/Qualifiers
     source          1..13049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             468..488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             498..524
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     protein_bind    complement(543..567)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    729..810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(766..785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    817..1041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1030..1047
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1079..2104)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(1079..2104)
                     /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(2114..2167)
                     /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    2191..2216
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    2223..3026
                     /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(3041..3059)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3069..3086)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3069..3085)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3078..3100)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3227..3682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3314..3333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3524..3545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3708..3812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3813..3881,3882..4673)
                     /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(4031..4050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4844..5432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5333..5352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5586..5603
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5720..5741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5756..5761,5762..5779,5780..5786)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5794..5810
                     /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     5799..5821
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5818..5834
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5818..5834
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5853..5873
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5855..5873
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    5892..6728
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        6759..7073
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    6767..6785
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6803..6821
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6838..6856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6874..6892
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6910..6928
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6945..6963
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6981..6999
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7046..7070
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7090..7114
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        7149..7167
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7217..7237
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7238..11338
                     /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             11396..11416
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gagcagcagt gaccacagag acagaagtga tctgggccaa ggccctgcca gcaggcacat
       61 ccgcccagcg ggccgagctg atcgccctga cccaggccct gaagatggcc gagggcaaga
      121 agctgaacgt gtacacagac tccagatatg ccttcgccac cgcacacatc cacggagaga
      181 tctacaggcg ccggggctgg ctgacctctg agggcaagga gatcaagaac aaggatgaga
      241 tcctggccct gctgaaggcc ctgtttctgc ccaagcggct gagcatcatc cactgtcctg
      301 gacaccagaa gggacactcc gccgaggcaa ggggcaatcg gatggccgac caggccgcca
      361 gaaaggctgc tattactgaa actcccgaca cttccactct gctgattgaa aactcctccc
      421 cttctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagtctccc aagaagaaga
      481 ggaaagtcgg ctctggccct gccgctaaga gagtgaagct ggacggaagc ggaggcgcgc
      541 cgacccagct ttcttgtaca aagtggttga tccggggatc ctctagtcag cgtttatctg
      601 cagaattcgc ccttgtttaa acgatcataa tcagccatac cacatttgta gaggttttac
      661 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
      721 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
      781 atttcacaaa taaagcattt ttttcactgc ctcgacccat agagcccacc gcatccccag
      841 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
      901 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
      961 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     1021 gctggcaact agaaggcaca gtcgaggctg atcagcgggt ttaaacgggc cctctagact
     1081 attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta
     1141 cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc
     1201 cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat
     1261 tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga
     1321 gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca
     1381 tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga
     1441 acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt
     1501 tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca
     1561 tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc
     1621 agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac
     1681 cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga
     1741 tcgcatccat ggcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt
     1801 cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt
     1861 ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat
     1921 aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc
     1981 ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga
     2041 cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt
     2101 tcatctgcag cctagggccg ggattctcct ccacgtcacc gcatgttaga agacttcctc
     2161 tgccctctcc gctgccagat ctctcgaggc cctgtgggag gaagagaaga ggtcagggat
     2221 ctgccccact gtggggtgga ggggacagat aaaagtaccc agaaccagag ccacattaac
     2281 cggccctggg aatataaggt ggtcccagct cggggacaca ggatccctgg aggcagcaaa
     2341 catgctgtcc tgaagtggac ataggggccc gggttggagg aagaagacta gctgagctct
     2401 cggacccctg gaagatgcca tgacaggggg ctggaagagc tagcacagac tagagaggta
     2461 aggggggtag gggagctgcc caaatgaaag gagtgagagg tgacccgaat ccacaggaga
     2521 acggggtgtc caggcaaaga aagcaagagg atggagaggt ggctaaagcc agggagacgg
     2581 ggtactttgg ggttgtccag aaaaacggtg atgatgcagg cctacaagaa ggggaggcgg
     2641 gacgcaaggg agacatccgt cggagaaggc catcctaaga aacgagagat ggcacaggcc
     2701 ccagaaggag aaggaaaagg gaacccagcg agtgaagacg gcatggggtt gggtgaggga
     2761 ggagagatgc ccggagagga cccagacacg gggaggatcc gctcagagga catcacgtgg
     2821 tgcagcgccg agaaggaagt gctccggaaa gagcatcctt gggcagcaac acagcagaga
     2881 gcaaggggaa gagggagtgg aggaagacgg aacctgaagg aggcggcagg gaaggatctg
     2941 ggccagccgt agaggtgacc caggccacaa gctgcagaca gaaagcggca caggcccagg
     3001 ggagagaatg ctggtcagag aaagcaggta cccaattcgc cctatagtga gtcgtattac
     3061 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3121 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3181 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3241 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3301 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3361 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3421 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3481 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3541 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3601 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3661 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3781 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3841 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3901 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3961 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4021 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4081 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4141 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4201 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4261 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4321 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4381 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4441 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4501 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4561 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4621 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4681 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4741 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4801 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4861 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4921 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4981 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5041 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5101 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5161 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5221 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5281 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5341 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5401 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5461 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5521 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5581 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5641 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5701 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5761 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5821 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     5881 agctggagct cagagcagag ccaggaaccc ctgtagggaa ggggcaggag agccaggggc
     5941 atgagatggt ggacgaggaa gggggacagg gaagcctgag cgcctctcct gggcttgcca
     6001 aggactcaaa cccagaagcc cagagcaggg ccttagggaa gcgggaccct gctctgggcg
     6061 gaggaatatg tcccagatag cactggggac tctttaagga aagaaggatg gagaaagaga
     6121 aagggagtag aggcggccac gacctggtga acacctagga cgcaccattc tcacaaaggg
     6181 agttttccac acggacaccc ccctcctcac cacagccctg ccaggacggg gctggctact
     6241 ggccttatct cacaggtaaa actgacgcac ggaggaacaa tataaattgg ggactagaaa
     6301 ggtgaagagc caaagttaga actcaggacc aacttattct gattttgttt ttccaaactg
     6361 cttctcctct tgggaagtgt aaggaagctg cagcaccagg atcagtgaaa cgcaccagac
     6421 agccgcgtca gagcagctca ggttctggga gagggtagcg cagggtggcc actgagaacc
     6481 gggcaggtca cgcatccccc ccttccctcc caccccctgc caagctctcc ctcccaggat
     6541 cctctctggc tccatcgtaa gcaaacctta gaggttctgg caaggagaga gatggctcca
     6601 ggaaatgggg gtgtgtcacc agataaggaa tctgcctaac aggaggtggg ggttagaccc
     6661 aatatcagga gactaggaag gaggaggcct aaggatgggg cttttctgtc accaatcctg
     6721 tccctagttt aattaaccct ctagagctta gtactgtcga gtttactccc tatcagtgat
     6781 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc
     6841 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     6901 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag
     6961 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
     7021 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
     7081 ttccgatcaa caagtttgta caaaaaagca ggctccgcgg ttttcaggtt ggaccggtgc
     7141 gcggccgcta atacgactca ctatagggag agccgccacc atgaaacgga cagccgacgg
     7201 aagcgagttc gagtcaccaa agaagaagcg gaaagtcgac aagaagtaca gcatcggcct
     7261 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     7321 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     7381 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     7441 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     7501 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     7561 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     7621 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     7681 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     7741 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     7801 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     7861 ggccatcctg tctgccagac tgagcaagag cagaaagctg gaaaatctga tcgcccagct
     7921 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     7981 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     8041 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     8101 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     8161 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     8221 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     8281 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     8341 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     8401 gctcgtgaag ctgaagagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     8461 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     8521 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     8581 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     8641 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     8701 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     8761 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     8821 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     8881 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     8941 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     9001 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     9061 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     9121 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     9181 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     9241 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     9301 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     9361 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     9421 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     9481 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     9541 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     9601 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     9661 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     9721 catcaaccgg ctgtccgact acgatgtgga cgctatcgtg cctcagagct ttctgaagga
     9781 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     9841 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     9901 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     9961 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
    10021 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
    10081 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
    10141 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
    10201 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
    10261 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
    10321 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
    10381 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
    10441 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
    10501 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
    10561 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
    10621 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
    10681 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
    10741 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
    10801 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
    10861 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
    10921 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
    10981 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
    11041 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
    11101 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
    11161 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
    11221 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
    11281 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggtgactc
    11341 cggcggaagc tctggtggca gcaagcggac cgccgacggc tctgaattcg agagccctaa
    11401 gaagaaaaga aaggtgagcg gaggctctag cggcggaagc accctgaaca ttgaagacga
    11461 gtatagactg catgaaacaa gcaaggaacc cgacgtgtcc ctgggctcca cctggctgtc
    11521 cgactttccc caggcctggg ccgagacagg aggaatgggc ctggccgtgc ggcaggcacc
    11581 cctgatcatc cctctgaagg ccacctctac acccgtgagc atcaagcagt accctatgtc
    11641 tcaggaggcc agactgggca tcaagcctca catccagagg ctgctggacc agggcatcct
    11701 ggtgccatgc cagagcccct ggaacacacc actgctgccc gtgaagaagc caggcaccaa
    11761 tgactataga cccgtgcagg atctgagaga ggtgaacaag agggtggagg atatccaccc
    11821 caccgtgccc aacccttaca atctgctgtc cggcctgccc ccttctcacc agtggtatac
    11881 agtgctggac ctgaaggatg ccttcttttg tctgagactg caccctacca gccagccact
    11941 gttcgccttt gagtggaggg accctgagat gggcatctct ggccagctga cctggacacg
    12001 cctgcctcag ggcttcaaga atagcccaac actgtttaac gaggccctgc accgcgacct
    12061 ggcagatttc cggatccagc acccagatct gatcctgctg cagtacgtgg acgatctgct
    12121 gctggccgcc accagcgagc tggattgcca gcagggaaca cgcgccctgc tgcagaccct
    12181 gggaaacctg ggatataggg catccgccaa gaaggcccag atctgtcaga agcaggtgaa
    12241 gtacctgggc tatctgctga aggagggcca gagatggctg acagaggcca ggaaggagac
    12301 agtgatgggc cagccaacac ccaagacccc aagacagctg agggagttcc tgggcaaagc
    12361 aggattttgc aggctgttca tcccaggatt cgcagagatg gcagcacctc tgtacccact
    12421 gaccaagccg ggcaccctgt ttaattgggg ccctgaccag cagaaggcct atcaggagat
    12481 caagcaggcc ctgctgacag caccagccct gggcctgcca gacctgacca agcctttcga
    12541 gctgtttgtg gatgagaagc agggctacgc caagggcgtg ctgacccaga agctgggacc
    12601 atggagacgg cccgtggcct atctgtccaa gaagctggac ccagtggcag caggatggcc
    12661 accatgcctg aggatggtgg cagcaatcgc cgtgctgaca aaggatgccg gcaagctgac
    12721 catgggacag ccactggtca tcctggcacc acacgcagtg gaggccctgg tgaagcagcc
    12781 tccagatcgc tggctgtcta acgcccggat gacacactac caggccctgc tgctggacac
    12841 cgatcgcgtg cagtttggcc ctgtggtggc cctgaatcca gccaccctgc tgcctctgcc
    12901 agaggagggc ctgcagcaca actgtctgga catcctggca gaggcacacg gaacaaggcc
    12961 agacctgacc gatcagcccc tgcctgacgc cgatcacaca tggtataccg atggaagctc
    13021 cctgctgcag gagggccaga ggaaggcag
//
