LOCUS       sequence-429369-       13035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible PE2 system for controllable prime editing; this plasmid is
            used to insert PE2 prime editor in one allele of AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13035)
  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 13035)
  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-iPE2-Hygro
FEATURES             Location/Qualifiers
     source          1..13035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             469..489
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             498..515
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(536..560)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    722..803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(759..778)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    810..1034
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1023..1040
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1072..2097)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(1072..2097)
                     /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(2107..2160)
                     /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    2184..2209
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    2216..3019
                     /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(3034..3052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3062..3079)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3062..3078)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3071..3093)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3220..3675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3307..3326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3517..3538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3701..3805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3806..3874,3875..4666)
                     /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(4024..4043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4837..5425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5326..5345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5579..5596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5713..5734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5749..5754,5755..5772,5773..5779)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5787..5803
                     /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     5792..5814
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5811..5827
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5811..5827
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5846..5866
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5848..5866
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    5885..6721
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        6752..7066
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    6760..6778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6796..6814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6831..6849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6867..6885
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6903..6921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6938..6956
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6974..6992
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7039..7063
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7083..7107
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        7142..7160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7210..7230
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7231..9744,9745..9747,9748..11331)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7231..11331
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 gcgggagctg cggtgaccac cgagaccgag gtaatctggg ctaaagccct gccagccggg
       61 acatccgctc agcgggctga actgatagca ctcacccagg ccctaaagat ggcagaaggt
      121 aagaagctaa atgtttatac tgatagccgt tatgcttttg ctactgccca tatccatgga
      181 gaaatataca gaaggcgtgg gtggctcaca tcagaaggca aagagatcaa aaataaagac
      241 gagatcttgg ccctactaaa agccctcttt ctgcccaaaa gacttagcat aatccattgt
      301 ccaggacatc aaaagggaca cagcgccgag gctagaggca accggatggc tgaccaagcg
      361 gcccgaaagg cagccatcac agagactcca gacacctcta ccctcctcat agaaaattca
      421 tcaccctctg gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag
      481 aggaaagtct aaccggtcat catcaccatc accattgagt ttggtgggcg cgccgaccca
      541 gctttcttgt acaaagtggt tgatccgggg atcctctagt cagcgtttat ctgcagaatt
      601 cgcccttgtt taaacgatca taatcagcca taccacattt gtagaggttt tacttgcttt
      661 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
      721 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
      781 aaataaagca tttttttcac tgcctcgacc catagagccc accgcatccc cagcatgcct
      841 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
      901 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
      961 agtggcacct tccagggtca aggaaggcac gggggaaggg caaacaacag atggctggca
     1021 actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag actattcctt
     1081 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
     1141 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
     1201 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
     1261 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg
     1321 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
     1381 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
     1441 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
     1501 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
     1561 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
     1621 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
     1681 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
     1741 catggcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa
     1801 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat
     1861 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
     1921 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
     1981 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
     2041 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatctg
     2101 cagcctaggg ccgggattct cctccacgtc accgcatgtt agaagacttc ctctgccctc
     2161 tccgctgcca gatctctcga ggccctgtgg gaggaagaga agaggtcagg gatctgcccc
     2221 actgtggggt ggaggggaca gataaaagta cccagaacca gagccacatt aaccggccct
     2281 gggaatataa ggtggtccca gctcggggac acaggatccc tggaggcagc aaacatgctg
     2341 tcctgaagtg gacatagggg cccgggttgg aggaagaaga ctagctgagc tctcggaccc
     2401 ctggaagatg ccatgacagg gggctggaag agctagcaca gactagagag gtaagggggg
     2461 taggggagct gcccaaatga aaggagtgag aggtgacccg aatccacagg agaacggggt
     2521 gtccaggcaa agaaagcaag aggatggaga ggtggctaaa gccagggaga cggggtactt
     2581 tggggttgtc cagaaaaacg gtgatgatgc aggcctacaa gaaggggagg cgggacgcaa
     2641 gggagacatc cgtcggagaa ggccatccta agaaacgaga gatggcacag gccccagaag
     2701 gagaaggaaa agggaaccca gcgagtgaag acggcatggg gttgggtgag ggaggagaga
     2761 tgcccggaga ggacccagac acggggagga tccgctcaga ggacatcacg tggtgcagcg
     2821 ccgagaagga agtgctccgg aaagagcatc cttgggcagc aacacagcag agagcaaggg
     2881 gaagagggag tggaggaaga cggaacctga aggaggcggc agggaaggat ctgggccagc
     2941 cgtagaggtg acccaggcca caagctgcag acagaaagcg gcacaggccc aggggagaga
     3001 atgctggtca gagaaagcag gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     3061 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3121 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3181 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     3241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3661 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4321 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4981 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5221 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5281 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5581 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5641 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5701 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     5761 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5821 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     5881 gctcagagca gagccaggaa cccctgtagg gaaggggcag gagagccagg ggcatgagat
     5941 ggtggacgag gaagggggac agggaagcct gagcgcctct cctgggcttg ccaaggactc
     6001 aaacccagaa gcccagagca gggccttagg gaagcgggac cctgctctgg gcggaggaat
     6061 atgtcccaga tagcactggg gactctttaa ggaaagaagg atggagaaag agaaagggag
     6121 tagaggcggc cacgacctgg tgaacaccta ggacgcacca ttctcacaaa gggagttttc
     6181 cacacggaca cccccctcct caccacagcc ctgccaggac ggggctggct actggcctta
     6241 tctcacaggt aaaactgacg cacggaggaa caatataaat tggggactag aaaggtgaag
     6301 agccaaagtt agaactcagg accaacttat tctgattttg tttttccaaa ctgcttctcc
     6361 tcttgggaag tgtaaggaag ctgcagcacc aggatcagtg aaacgcacca gacagccgcg
     6421 tcagagcagc tcaggttctg ggagagggta gcgcagggtg gccactgaga accgggcagg
     6481 tcacgcatcc cccccttccc tcccaccccc tgccaagctc tccctcccag gatcctctct
     6541 ggctccatcg taagcaaacc ttagaggttc tggcaaggag agagatggct ccaggaaatg
     6601 ggggtgtgtc accagataag gaatctgcct aacaggaggt gggggttaga cccaatatca
     6661 ggagactagg aaggaggagg cctaaggatg gggcttttct gtcaccaatc ctgtccctag
     6721 tttaattaac cctctagagc ttagtactgt cgagtttact ccctatcagt gatagagaac
     6781 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
     6841 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
     6901 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
     6961 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
     7021 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattccgat
     7081 caacaagttt gtacaaaaaa gcaggctccg cggttttcag gttggaccgg tgcgcggccg
     7141 ctaatacgac tcactatagg gagagccgcc accatgaaac ggacagccga cggaagcgag
     7201 ttcgagtcac caaagaagaa gcggaaagtc gacaagaagt acagcatcgg cctggacatc
     7261 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     7321 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     7381 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     7441 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     7501 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     7561 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     7621 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     7681 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     7741 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     7801 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     7861 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     7921 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     7981 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     8041 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     8101 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     8161 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     8221 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     8281 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     8341 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     8401 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     8461 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     8521 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     8581 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     8641 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     8701 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     8761 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     8821 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     8881 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     8941 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     9001 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     9061 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     9121 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     9181 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     9241 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     9301 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     9361 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     9421 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     9481 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     9541 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     9601 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     9661 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     9721 cggctgtccg actacgatgt ggacgctatc gtgcctcaga gctttctgaa ggacgactcc
     9781 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     9841 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     9901 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     9961 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
    10021 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
    10081 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
    10141 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
    10201 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
    10261 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
    10321 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
    10381 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
    10441 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
    10501 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
    10561 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
    10621 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
    10681 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
    10741 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
    10801 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
    10861 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
    10921 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
    10981 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
    11041 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
    11101 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
    11161 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
    11221 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
    11281 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggtga ctctggagga
    11341 tctagcggag gatcctctgg cagcgagaca ccaggaacaa gcgagtcagc aacaccagag
    11401 agcagtggcg gcagcagcgg cggcagcagc accctaaata tagaagatga gtatcggcta
    11461 catgagacct caaaagagcc agatgtttct ctagggtcca catggctgtc tgattttcct
    11521 caggcctggg cggaaaccgg gggcatggga ctggcagttc gccaagctcc tctgatcata
    11581 cctctgaaag caacctctac ccccgtgtcc ataaaacaat accccatgtc acaagaagcc
    11641 agactgggga tcaagcccca catacagaga ctgttggacc agggaatact ggtaccctgc
    11701 cagtccccct ggaacacgcc cctgctaccc gttaagaaac cagggactaa tgattatagg
    11761 cctgtccagg atctgagaga agtcaacaag cgggtggaag acatccaccc caccgtgccc
    11821 aacccttaca acctcttgag cgggctccca ccgtcccacc agtggtacac tgtgcttgat
    11881 ttaaaggatg cctttttctg cctgagactc caccccacca gtcagcctct cttcgccttt
    11941 gagtggagag atccagagat gggaatctca ggacaattga cctggaccag actcccacag
    12001 ggtttcaaaa acagtcccac cctgtttaat gaggcactgc acagagacct agcagacttc
    12061 cggatccagc acccagactt gatcctgcta cagtacgtgg atgacttact gctggccgcc
    12121 acttctgagc tagactgcca acaaggtact cgggccctgt tacaaaccct agggaacctc
    12181 gggtatcggg cctcggccaa gaaagcccaa atttgccaga aacaggtcaa gtatctgggg
    12241 tatcttctaa aagagggtca gagatggctg actgaggcca gaaaagagac tgtgatgggg
    12301 cagcctactc cgaagacccc tcgacaacta agggagttcc tagggaaggc aggcttctgt
    12361 cgcctcttca tccctgggtt tgcagaaatg gcagcccccc tgtaccctct caccaaaccg
    12421 gggactctgt ttaattgggg cccagaccaa caaaaggcct atcaagaaat caagcaagct
    12481 cttctaactg ccccagccct ggggttgcca gatttgacta agccctttga actctttgtc
    12541 gacgagaagc agggctacgc caaaggtgtc ctaacgcaaa aactgggacc ttggcgtcgg
    12601 ccggtggcct acctgtccaa aaagctagac ccagtagcag ctgggtggcc cccttgccta
    12661 cggatggtag cagccattgc cgtactgaca aaggatgcag gcaagctaac catgggacag
    12721 ccactagtca ttctggcccc ccatgcagta gaggcactag tcaaacaacc ccccgaccgc
    12781 tggctttcca acgcccggat gactcactat caggccttgc ttttggacac ggaccgggtc
    12841 cagttcggac cggtggtagc cctgaacccg gctacgctgc tcccactgcc tgaggaaggg
    12901 ctgcaacaca actgccttga tatcctggcc gaagcccacg gaacccgacc cgacctaacg
    12961 gaccagccgc tcccagacgc cgaccacacc tggtacacgg atggaagcag tctcttacaa
    13021 gagggacagc gtaag
//
