LOCUS       sequence-367737-       13134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13134)
  AUTHORS   Velimirovic M, Zanetti LC, Shen MW, Fife JD, Lin L, Cha M, Akinci E,
            Barnum D, Yu T, Sherwood RI
  TITLE     Peptide fusion improves prime editing efficiency.
  JOURNAL   Nat Commun. 2022 Jun 18;13(1):3512. doi: 10.1038/s41467-022-31270-y.
  PUBMED    35717416
REFERENCE   2  (bases 1 to 13134)
  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: "2022-06-18"; volume: "13(1):3512. doi"; pages: " 
            10.1038/s41467-022-31270-y"
COMMENT     Sequence Label: p2T_CAG_SpCas9PE2_5plinker_BlastR
FEATURES             Location/Qualifiers
     source          1..13134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(679..690,691..853)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     regulatory      878..887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(893..926)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        941..1320
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1322..1599
                     /label=chicken beta-actin promoter
     intron          1600..2616
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2540..2562
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2624..2643
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2858..2878
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2903..5416,5417..5419,5420..7003)
                     /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             2903..7003
                     /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"
     CDS             9176..9196
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9205..9222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9245..9262)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9251..9475
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    9546..9680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9570..9589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9624..9643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9838..10236)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(10255..10302)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(10350..10679)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10364..10499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10418..10437)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     10664..10684
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(10694..10881,10882..10893)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(10950..10970)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10950..10968)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10989..11005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10989..11005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11002..11024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11013..11029
                     /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)."
     promoter        complement(join(11037..11043,11044..11061,11062..11067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11082..11103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11220..11237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11391..11979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11471..11490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12150..12941,12942..13010))
                     /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     12773..12792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13011..13115)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 ccggcatatg gttcttgaca gaggtgtaaa aagtactcaa aaattttact caagtgaaag
      721 tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta
      781 cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattaa ggaagtaaaa
      841 gtaaaagcaa gaaagatcga tctcgaagga tctggaggcc accatggtgt cgataacttc
      901 gtatagcata cattatacga agttatcgat aagcttgctc gacattgatt attgactagt
      961 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1021 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1081 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1141 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1201 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1261 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1321 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     1381 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     1441 gggcgcgcgc crggsggggs ggggsggggs grggggsggg gsggggcgag gcggagaggt
     1501 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     1561 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     1621 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     1681 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
     1741 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
     1801 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
     1861 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
     1921 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1981 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     2041 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     2101 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     2161 ggcggggggt ggcggcaggk gggggtgccg ggcggggcgg ggccgcctcg ggccggggrg
     2221 ggctcggggg rggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     2281 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     2341 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     2401 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     2461 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     2521 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     2581 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     2641 gtgctgtctc atcattttgg caaagaattc ctcgagcggc cgccagtgtg atggatatct
     2701 gcagaattcc agcacactgg cggccgttac tagtggatcc accggtacca tggctagcgg
     2761 aggatctagc ggaggctcct ctggctctga gacacctggc acaagcgaga gcgcaacacc
     2821 tgaaagcagc gggggcagca gcggggggtc atgtacacca aagaagaagc ggaaggtcgg
     2881 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     2941 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     3001 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     3061 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     3121 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     3181 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     3241 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     3301 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     3361 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     3421 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     3481 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     3541 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     3601 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     3661 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     3721 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     3781 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     3841 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     3901 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     3961 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     4021 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     4081 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     4141 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     4201 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     4261 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     4321 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     4381 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     4441 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     4501 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     4561 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     4621 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     4681 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     4741 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     4801 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     4861 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     4921 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     4981 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     5041 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     5101 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     5161 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     5221 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     5281 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     5341 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     5401 cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     5461 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     5521 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     5581 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     5641 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     5701 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     5761 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     5821 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     5881 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     5941 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     6001 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     6061 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     6121 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     6181 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     6241 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     6301 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     6361 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     6421 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     6481 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     6541 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     6601 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     6661 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     6721 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     6781 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     6841 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     6901 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     6961 cctgtacgag acacggatcg acctgtctca gctgggaggt gactctggag gatctagcgg
     7021 aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag agagcagtgg
     7081 cggcagcagc ggcggcagca gcaccctaaa tatagaagat gagtatcggc tacatgagac
     7141 ctcaaaagag ccagatgttt ctctagggtc cacatggctg tctgattttc ctcaggcctg
     7201 ggcggaaacc gggggcatgg gactggcagt tcgccaagct cctctgatca tacctctgaa
     7261 agcaacctct acccccgtgt ccataaaaca ataccccatg tcacaagaag ccagactggg
     7321 gatcaagccc cacatacaga gactgttgga ccagggaata ctggtaccct gccagtcccc
     7381 ctggaacacg cccctgctac ccgttaagaa accagggact aatgattata ggcctgtcca
     7441 ggatctgaga gaagtcaaca agcgggtgga agacatccac cccaccgtgc ccaaccctta
     7501 caacctcttg agcgggctcc caccgtccca ccagtggtac actgtgcttg atttaaagga
     7561 tgcctttttc tgcctgagac tccaccccac cagtcagcct ctcttcgcct ttgagtggag
     7621 agatccagag atgggaatct caggacaatt gacctggacc agactcccac agggtttcaa
     7681 aaacagtccc accctgttta atgaggcact gcacagagac ctagcagact tccggatcca
     7741 gcacccagac ttgatcctgc tacagtacgt ggatgactta ctgctggccg ccacttctga
     7801 gctagactgc caacaaggta ctcgggccct gttacaaacc ctagggaacc tcgggtatcg
     7861 ggcctcggcc aagaaagccc aaatttgcca gaaacaggtc aagtatctgg ggtatcttct
     7921 aaaagagggt cagagatggc tgactgaggc cagaaaagag actgtgatgg ggcagcctac
     7981 tccgaagacc cctcgacaac taagggagtt cctagggaag gcaggcttct gtcgcctctt
     8041 catccctggg tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac cggggactct
     8101 gtttaattgg ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag ctcttctaac
     8161 tgccccagcc ctggggttgc cagatttgac taagcccttt gaactctttg tcgacgagaa
     8221 gcagggctac gccaaaggtg tcctaacgca aaaactggga ccttggcgtc ggccggtggc
     8281 ctacctgtcc aaaaagctag acccagtagc agctgggtgg cccccttgcc tacggatggt
     8341 agcagccatt gccgtactga caaaggatgc aggcaagcta accatgggac agccactagt
     8401 cattctggcc ccccatgcag tagaggcact agtcaaacaa ccccccgacc gctggctttc
     8461 caacgcccgg atgactcact atcaggcctt gcttttggac acggaccggg tccagttcgg
     8521 accggtggta gccctgaacc cggctacgct gctcccactg cctgaggaag ggctgcaaca
     8581 caactgcctt gatatcctgg ccgaagccca cggaacccga cccgacctaa cggaccagcc
     8641 gctcccagac gccgaccaca cctggtacac ggatggaagc agtctcttac aagagggaca
     8701 gcgtaaggcg ggagctgcgg tgaccaccga gaccgaggta atctgggcta aagccctgcc
     8761 agccgggaca tccgctcagc gggctgaact gatagcactc acccaggccc taaagatggc
     8821 agaaggtaag aagctaaatg tttatactga tagccgttat gcttttgcta ctgcccatat
     8881 ccatggagaa atatacagaa ggcgtgggtg gctcacatca gaaggcaaag agatcaaaaa
     8941 taaagacgag atcttggccc tactaaaagc cctctttctg cccaaaagac ttagcataat
     9001 ccattgtcca ggacatcaaa agggacacag cgccgaggct agaggcaacc ggatggctga
     9061 ccaagcggcc cgaaaggcag ccatcacaga gactccagac acctctaccc tcctcataga
     9121 aaattcatca ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa
     9181 gaagaagagg aaagtctaac cggtcatcat caccatcacc attgagttta aacccgctga
     9241 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     9301 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     9361 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     9421 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     9481 gaggcggaaa gaacaagatc tagcggccgc caccgcggtg gagctcgaat taattcatcg
     9541 atgatgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     9601 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     9661 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     9721 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     9781 gcggtggaat cgaaatctcg tagcacgtgt cagtcctgct cctcggccac gaagtgctta
     9841 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     9901 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     9961 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
    10021 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
    10081 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
    10141 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
    10201 gagggtggat tcttcttgag acaaaggctt ggccatggtt tagttcctca ccttgtcgta
    10261 ttatactatg ccgatatact atgccgatga ttaattgtca acacgtgctg atcagatccg
    10321 aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
    10381 cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa
    10441 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
    10501 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
    10561 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
    10621 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
    10681 ttccacagaa ttcaagtgat ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt
    10741 aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg
    10801 atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat
    10861 caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc ttcagcaagc
    10921 ccagcagatc cgagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
    10981 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    11041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    11101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    11161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    11221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    11281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    11341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    11401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    11461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    11521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    11581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    11641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    11701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    11761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    11821 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    11881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    11941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    12001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    12061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    12121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    12181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    12241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    12301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    12361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    12421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    12481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    12541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    12601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    12661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    12721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    12781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    12841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    12901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    12961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    13021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    13081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtg
//
