LOCUS       sequence-442579-       16266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HDR donor plasmid for PE7 knockin at AAVS1 locus. 
            Left_homology_arm-splicing_acceptor-T2A-NeoR-bGHpA-pEF1-alpha-PE7-IR
            ES2-EGFP-WPRE-beta-globinpA-right_homology_arm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16266)
  AUTHORS   Yan J, Oyler-Castrillo P, Ravisankar P, Ward CC, Levesque S, Jing Y,
            Simpson D, Zhao A, Li H, Yan W, Goudy L, Schmidt R, Solley SC, 
            Gilbert LA, Chan MM, Bauer DE, Marson A, Parsons LR, Adamson B
  TITLE     Improving prime editing with an endogenous small RNA-binding 
            protein.
  JOURNAL   Nature. 2024 Apr 3. doi: 10.1038/s41586-024-07259-6.
  PUBMED    38570691
REFERENCE   2  (bases 1 to 16266)
  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: "Nature. 
            2024 Apr 3. doi: 10.1038/s41586-024-07259-6."
COMMENT     Sequence Label: PE7_AAVS1_knockin_HDR_donor
FEATURES             Location/Qualifiers
     source          1..16266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..804
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    823..848
                     /label=SA
                     /note="splice acceptor site"
     CDS             872..925
                     /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"
     CDS             935..1735
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GTMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(995..1014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1605..1624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1755..1772)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1761..1985
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2088..2299
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2300..3238
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3195..3215
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        3260..3278
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3328..3348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3349..7449
                     /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             7507..7527
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10306..10326
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10336..10362
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             10371..10388
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    join(10441..11015,11016..11018,11019..11027)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(10608..10625)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     10835..10854
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(11028..11030,11031..11033,11034..11747)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(11028..11030,11031..11033,11034..11744)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(11334..11353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11681..11702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(11767..11800)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    11856..12444
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11909..11929)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12327..12338)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    12476..12531
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(12530..12549)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    12749..13278
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(13425..13443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13450..13467)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13450..13466)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13459..13481)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      13608..14063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13695..13714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13905..13926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        14089..14193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14194..14262,14263..15054)
                     /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(14412..14431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      15225..15813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15714..15733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     15967..15984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    16101..16122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(16137..16142,16143..16160,16161..16167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16175..16191
                     /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     16180..16202
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     16199..16215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16199..16215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        16236..16254
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgctttctct gaccagcatt ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt
       61 ggcctgggtc acctctacgg ctggcccaga tccttccctg ccgcctcctt caggttccgt
      121 cttcctccac tccctcttcc ccttgctctc tgctgtgttg ctgcccaagg atgctctttc
      181 cggagcactt ccttctcggc gctgcaccac gtgatgtcct ctgagcggat cctccccgtg
      241 tctgggtcct ctccgggcat ctctcctccc tcacccaacc ccatgccgtc ttcactcgct
      301 gggttccctt ttccttctcc ttctggggcc tgtgccatct ctcgtttctt aggatggcct
      361 tctccgacgg atgtctccct tgcgtcccgc ctccccttct tgtaggcctg catcatcacc
      421 gtttttctgg acaaccccaa agtaccccgt ctccctggct ttagccacct ctccatcctc
      481 ttgctttctt tgcctggaca ccccgttctc ctgtggattc gggtcacctc tcactccttt
      541 catttgggca gctcccctac cccccttacc tctctagtct gtgctagctc ttccagcccc
      601 ctgtcatggc atcttccagg ggtccgagag ctcagctagt cttcttcctc caacccgggc
      661 ccctatgtcc acttcaggac agcatgtttg ctgcctccag ggatcctgtg tccccgagct
      721 gggaccacct tatattccca gggccggtta atgtggctct ggttctgggt acttttatct
      781 gtcccctcca ccccacagtg gggccactag ggacagaagc ttctgacctc ttctcttcct
      841 cccacagggc ctcgagagat ctggcagcgg agagggcaga ggaagtcttc taacatgcgg
      901 tgacgtggag gagaatcccg gccctaggct cgagggtacc atgattgaac aagatggatt
      961 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1021 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1081 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1141 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1201 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1261 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1321 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1381 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1441 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1501 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1561 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1621 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1681 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagttta
     1741 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1801 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1861 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1921 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1981 tatggactag tattaattaa atcactcacg cgtctaggat ccatgccgat agcgttggtt
     2041 gagtggataa ccgtattacc gccaagctta tgcatgtgcc cgtcagtggg cagagcgcac
     2101 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
     2161 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2221 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2281 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
     2341 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga
     2401 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
     2461 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg
     2521 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
     2581 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
     2641 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
     2701 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     2761 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     2821 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     2881 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     2941 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     3001 gtaccgggcg ccgtccaggc acctcgatta gttctcgcgc ttttggagta cgtcgtcttt
     3061 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     3121 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     3181 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     3241 gtcgtgagct agcggccgct aatacgactc actataggga gagccgccac catgaaacgg
     3301 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtcga caagaagtac
     3361 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     3421 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     3481 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     3541 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     3601 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     3661 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     3721 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     3781 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     3841 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     3901 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     3961 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagaaagct ggaaaatctg
     4021 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     4081 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     4141 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     4201 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     4261 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     4321 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     4381 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     4441 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     4501 accgaggaac tgctcgtgaa gctgaagaga gaggacctgc tgcggaagca gcggaccttc
     4561 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     4621 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     4681 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     4741 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     4801 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     4861 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     4921 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     4981 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     5041 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     5101 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     5161 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     5221 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     5281 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     5341 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     5401 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     5461 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     5521 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     5581 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     5641 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     5701 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     5761 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     5821 caggaactgg acatcaaccg gctgtccgac tacgatgtgg acgctatcgt gcctcagagc
     5881 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     5941 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     6001 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     6061 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     6121 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     6181 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     6241 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     6301 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     6361 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     6421 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     6481 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     6541 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     6601 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     6661 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     6721 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     6781 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     6841 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     6901 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     6961 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     7021 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     7081 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     7141 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     7201 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     7261 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     7321 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     7381 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     7441 ggaggtgact ccggcggaag ctctggtggc agcaagcgga ccgccgacgg ctctgaattc
     7501 gagagcccta agaagaaaag aaaggtgagc ggaggctcta gcggcggaag caccctgaac
     7561 attgaagacg agtatagact gcatgaaaca agcaaggaac ccgacgtgtc cctgggctcc
     7621 acctggctgt ccgactttcc ccaggcctgg gccgagacag gaggaatggg cctggccgtg
     7681 cggcaggcac ccctgatcat ccctctgaag gccacctcta cacccgtgag catcaagcag
     7741 taccctatgt ctcaggaggc cagactgggc atcaagcctc acatccagag gctgctggac
     7801 cagggcatcc tggtgccatg ccagagcccc tggaacacac cactgctgcc cgtgaagaag
     7861 ccaggcacca atgactatag acccgtgcag gatctgagag aggtgaacaa gagggtggag
     7921 gatatccacc ccaccgtgcc caacccttac aatctgctgt ccggcctgcc cccttctcac
     7981 cagtggtata cagtgctgga cctgaaggat gccttctttt gtctgagact gcaccctacc
     8041 agccagccac tgttcgcctt tgagtggagg gaccctgaga tgggcatctc tggccagctg
     8101 acctggacac gcctgcctca gggcttcaag aatagcccaa cactgtttaa cgaggccctg
     8161 caccgcgacc tggcagattt ccggatccag cacccagatc tgatcctgct gcagtacgtg
     8221 gacgatctgc tgctggccgc caccagcgag ctggattgcc agcagggaac acgcgccctg
     8281 ctgcagaccc tgggaaacct gggatatagg gcatccgcca agaaggccca gatctgtcag
     8341 aagcaggtga agtacctggg ctatctgctg aaggagggcc agagatggct gacagaggcc
     8401 aggaaggaga cagtgatggg ccagccaaca cccaagaccc caagacagct gagggagttc
     8461 ctgggcaaag caggattttg caggctgttc atcccaggat tcgcagagat ggcagcacct
     8521 ctgtacccac tgaccaagcc gggcaccctg tttaattggg gccctgacca gcagaaggcc
     8581 tatcaggaga tcaagcaggc cctgctgaca gcaccagccc tgggcctgcc agacctgacc
     8641 aagcctttcg agctgtttgt ggatgagaag cagggctacg ccaagggcgt gctgacccag
     8701 aagctgggac catggagacg gcccgtggcc tatctgtcca agaagctgga cccagtggca
     8761 gcaggatggc caccatgcct gaggatggtg gcagcaatcg ccgtgctgac aaaggatgcc
     8821 ggcaagctga ccatgggaca gccactggtc atcctggcac cacacgcagt ggaggccctg
     8881 gtgaagcagc ctccagatcg ctggctgtct aacgcccgga tgacacacta ccaggccctg
     8941 ctgctggaca ccgatcgcgt gcagtttggc cctgtggtgg ccctgaatcc agccaccctg
     9001 ctgcctctgc cagaggaggg cctgcagcac aactgtctgg acatcctggc agaggcacac
     9061 ggaacaaggc cagacctgac cgatcagccc ctgcctgacg ccgatcacac atggtatacc
     9121 gatggaagct ccctgctgca ggagggccag aggaaggcag gagcagcagt gaccacagag
     9181 acagaagtga tctgggccaa ggccctgcca gcaggcacat ccgcccagcg ggccgagctg
     9241 atcgccctga cccaggccct gaagatggcc gagggcaaga agctgaacgt gtacacagac
     9301 tccagatatg ccttcgccac cgcacacatc cacggagaga tctacaggcg ccggggctgg
     9361 ctgacctctg agggcaagga gatcaagaac aaggatgaga tcctggccct gctgaaggcc
     9421 ctgtttctgc ccaagcggct gagcatcatc cactgtcctg gacaccagaa gggacactcc
     9481 gccgaggcaa ggggcaatcg gatggccgac caggccgcca gaaaggctgc tattactgaa
     9541 actcccgaca cttccactct gctgattgaa aactcctccc cttccggagg atctagcgga
     9601 ggctcctctg gctctgagac acctggcaca agcgagagcg caacacctga aagcagcggg
     9661 ggcagcagcg gggggtcaat ggctgaaaat ggtgataatg aaaagatggc tgccctggag
     9721 gccaaaatct gtcatcaaat tgagtattat tttggcgact tcaatttgcc acgggacaag
     9781 tttctaaagg aacagataaa actggatgaa ggctgggtac ctttggagat aatgataaaa
     9841 ttcaacaggt tgaaccgtct aacaacagac tttaatgtaa ttgtggaagc attgagcaaa
     9901 tccaaggcag aactcatgga aatcagtgaa gataaaacta aaatcagaag gtctccaagc
     9961 aaacccctac ctgaagtgac tgatgagtat aaaaatgatg taaaaaacag atctgtttat
    10021 attaaaggct tcccaactga tgcaactctt gatgacataa aagaatggtt agaagataaa
    10081 ggtcaagtac taaatattca gatgagaaga acattgcata aagcatttaa gggatcaatt
    10141 tttgttgtgt ttgatagcat tgaatctgct aagaaatttg tagagacccc tggccagaag
    10201 tacaaagaaa cagacctgct aatacttttc aaggacgatt actttgccaa aaaaaatgaa
    10261 tctggcggct caaaaagaac cgccgacggc agcgaattcg agtctcccaa gaagaagagg
    10321 aaagtcggct ctggccctgc cgctaagaga gtgaagctgg actaaccggt catcatcacc
    10381 atcaccattg agtttgctcg gccaattctg cagtcgacgg taccgcgggc ccgggatccg
    10441 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
    10501 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
    10561 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
    10621 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
    10681 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
    10741 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
    10801 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
    10861 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggta
    10921 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
    10981 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
    11041 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
    11101 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
    11161 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
    11221 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
    11281 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
    11341 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
    11401 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
    11461 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
    11521 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
    11581 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
    11641 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
    11701 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagaa ttcgtcgagg
    11761 gacctaataa cttcgtatag catacattat acgaagttat acatgtttaa gggttccggt
    11821 tccactaggt acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
    11881 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
    11941 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
    12001 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
    12061 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
    12121 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
    12181 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
    12241 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
    12301 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
    12361 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
    12421 tctccctttg ggccgcctcc ccgcatgaag ccccttgagc atctgacttc tggctaataa
    12481 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
    12541 acatatggga gggcaaatca taagtcgaca gtactaagct tgattggtga cagaaaagcc
    12601 ccatccttag gcctcctcct tcctagtctc ctgatattgg gtctaacccc cacctcctgt
    12661 taggcagatt ccttatctgg tgacacaccc ccatttcctg gagccatctc tctccttgcc
    12721 agaacctcta aggtttgctt acgatggagc cagagaggat cctgggaggg agagcttggc
    12781 agggggtggg agggaagggg gggatgcgtg acctgcccgg ttctcagtgg ccaccctgcg
    12841 ctaccctctc ccagaacctg agctgctctg acgcggctgt ctggtgcgtt tcactgatcc
    12901 tggtgctgca gcttccttac acttcccaag aggagaagca gtttggaaaa acaaaatcag
    12961 aataagttgg tcctgagttc taactttggc tcttcacctt tctagtcccc aatttatatt
    13021 gttcctccgt gcgtcagttt tacctgtgag ataaggccag tagccagccc cgtcctggca
    13081 gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga atggtgcgtc
    13141 ctaggtgttc accaggtcgt ggccgcctct actccctttc tctttctcca tccttctttc
    13201 cttaaagagt ccccagtgct atctgggaca tattcctccg cccagagcag ggtcccgctt
    13261 ccctaaggcc ctgctctggg cttctgggtt tgagtccttg gcaagcccag gagaggcgct
    13321 caggcttccc tgtccccctt cctcgtccac catctcatgc ccctggctct cctgcccctt
    13381 ccctacaggg gttcctggct ctgctctagc gatcgccaat tcgccctata gtgagtcgta
    13441 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
    13501 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
    13561 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
    13621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
    13681 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
    13741 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
    13801 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
    13861 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
    13921 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
    13981 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
    14041 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
    14101 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
    14161 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
    14221 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
    14281 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
    14341 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
    14401 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
    14461 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
    14521 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
    14581 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    14641 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    14701 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    14761 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    14821 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    14881 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    14941 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    15001 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    15061 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    15121 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    15181 ttccactgag cgtcaaaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    15241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    15301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    15361 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    15421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgtcag tggcgataag
    15481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    15541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    15601 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    15661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    15721 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    15781 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    15841 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    15901 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    15961 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    16021 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    16081 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    16141 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    16201 ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa
    16261 aagctg
//
