LOCUS       sequence-386782-       10125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SpCas9 prime editor 2 with P2A-p53DD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10125)
  AUTHORS   Li M, Zhong A, Wu Y, Sidharta M, Beaury M, Zhao X, Studer L, Zhou T
  TITLE     Transient inhibition of p53 enhances prime editing and cytosine 
            base-editing efficiencies in human pluripotent stem cells.
  JOURNAL   Nat Commun. 2022 Oct 27;13(1):6354. doi: 10.1038/s41467-022-34045-7.
  PUBMED    36302757
REFERENCE   2  (bases 1 to 10125)
  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: "2022-10-27"; volume: "13(1):6354. doi"; pages: " 
            10.1038/s41467-022-34045-7"
COMMENT     Sequence Label: pCMV-PE2-p53DD
FEATURES             Location/Qualifiers
     source          1..10125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        39..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     664..683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        664..682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             732..752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(753..3266,3267..3269,3270..4853)
                     /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             753..4853
                     /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             7026..7046
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7056..7112
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7427..7444
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7467..7484)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7473..7697
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7768..7784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7768..7784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7781..7803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7792..7808
                     /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(7816..7822,7823..7840,7841..7846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7861..7882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7999..8016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8170..8758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8250..8269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8929..9720,9721..9789))
                     /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     9552..9571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9790..9894)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9973..9992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctaga gatccgcggc
      661 cgctaatacg actcactata gggagagccg ccaccatgaa acggacagcc gacggaagcg
      721 agttcgagtc accaaagaag aagcggaaag tcgacaagaa gtacagcatc ggcctggaca
      781 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga
      841 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc
      901 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa
      961 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg
     1021 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata
     1081 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga
     1141 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc
     1201 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg
     1261 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga
     1321 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca
     1381 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg
     1441 gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca
     1501 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct
     1561 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc
     1621 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg
     1681 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg
     1741 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt
     1801 tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag
     1861 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg
     1921 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc
     1981 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc
     2041 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact
     2101 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg
     2161 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct
     2221 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc
     2281 acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga
     2341 ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc
     2401 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga
     2461 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc
     2521 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg
     2581 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga
     2641 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc
     2701 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc
     2761 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca
     2821 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag
     2881 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc
     2941 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga
     3001 tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc
     3061 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc
     3121 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc
     3181 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca
     3241 accggctgtc cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact
     3301 ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc
     3361 cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc
     3421 tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac
     3481 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg
     3541 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc
     3601 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc
     3661 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg
     3721 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg
     3781 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca
     3841 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta
     3901 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg
     3961 agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc
     4021 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc
     4081 tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt
     4141 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
     4201 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa
     4261 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
     4321 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
     4381 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
     4441 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
     4501 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
     4561 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
     4621 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
     4681 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
     4741 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga
     4801 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggt gactctggag
     4861 gatctagcgg aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag
     4921 agagcagtgg cggcagcagc ggcggcagca gcaccctaaa tatagaagat gagtatcggc
     4981 tacatgagac ctcaaaagag ccagatgttt ctctagggtc cacatggctg tctgattttc
     5041 ctcaggcctg ggcggaaacc gggggcatgg gactggcagt tcgccaagct cctctgatca
     5101 tacctctgaa agcaacctct acccccgtgt ccataaaaca ataccccatg tcacaagaag
     5161 ccagactggg gatcaagccc cacatacaga gactgttgga ccagggaata ctggtaccct
     5221 gccagtcccc ctggaacacg cccctgctac ccgttaagaa accagggact aatgattata
     5281 ggcctgtcca ggatctgaga gaagtcaaca agcgggtgga agacatccac cccaccgtgc
     5341 ccaaccctta caacctcttg agcgggctcc caccgtccca ccagtggtac actgtgcttg
     5401 atttaaagga tgcctttttc tgcctgagac tccaccccac cagtcagcct ctcttcgcct
     5461 ttgagtggag agatccagag atgggaatct caggacaatt gacctggacc agactcccac
     5521 agggtttcaa aaacagtccc accctgttta atgaggcact gcacagagac ctagcagact
     5581 tccggatcca gcacccagac ttgatcctgc tacagtacgt ggatgactta ctgctggccg
     5641 ccacttctga gctagactgc caacaaggta ctcgggccct gttacaaacc ctagggaacc
     5701 tcgggtatcg ggcctcggcc aagaaagccc aaatttgcca gaaacaggtc aagtatctgg
     5761 ggtatcttct aaaagagggt cagagatggc tgactgaggc cagaaaagag actgtgatgg
     5821 ggcagcctac tccgaagacc cctcgacaac taagggagtt cctagggaag gcaggcttct
     5881 gtcgcctctt catccctggg tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac
     5941 cggggactct gtttaattgg ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag
     6001 ctcttctaac tgccccagcc ctggggttgc cagatttgac taagcccttt gaactctttg
     6061 tcgacgagaa gcagggctac gccaaaggtg tcctaacgca aaaactggga ccttggcgtc
     6121 ggccggtggc ctacctgtcc aaaaagctag acccagtagc agctgggtgg cccccttgcc
     6181 tacggatggt agcagccatt gccgtactga caaaggatgc aggcaagcta accatgggac
     6241 agccactagt cattctggcc ccccatgcag tagaggcact agtcaaacaa ccccccgacc
     6301 gctggctttc caacgcccgg atgactcact atcaggcctt gcttttggac acggaccggg
     6361 tccagttcgg accggtggta gccctgaacc cggctacgct gctcccactg cctgaggaag
     6421 ggctgcaaca caactgcctt gatatcctgg ccgaagccca cggaacccga cccgacctaa
     6481 cggaccagcc gctcccagac gccgaccaca cctggtacac ggatggaagc agtctcttac
     6541 aagagggaca gcgtaaggcg ggagctgcgg tgaccaccga gaccgaggta atctgggcta
     6601 aagccctgcc agccgggaca tccgctcagc gggctgaact gatagcactc acccaggccc
     6661 taaagatggc agaaggtaag aagctaaatg tttatactga tagccgttat gcttttgcta
     6721 ctgcccatat ccatggagaa atatacagaa ggcgtgggtg gctcacatca gaaggcaaag
     6781 agatcaaaaa taaagacgag atcttggccc tactaaaagc cctctttctg cccaaaagac
     6841 ttagcataat ccattgtcca ggacatcaaa agggacacag cgccgaggct agaggcaacc
     6901 ggatggctga ccaagcggcc cgaaaggcag ccatcacaga gactccagac acctctaccc
     6961 tcctcataga aaattcatca ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat
     7021 tcgagcccaa gaagaagagg aaagtcggaa gcggagctac taacttcagc ctgctgaagc
     7081 aggctggaga cgtggaggag aaccctggac ctatgactgc catggaggag tcacagtcgg
     7141 atatcagcct caagagagcg ctgcccacct gcacaagcgc ctctcccccg caaaagaaaa
     7201 aaccacttga tggagagtat ttcaccctca agatccgcgg gcgtaaacgc ttcgagatgt
     7261 tccgggagct gaatgaggcc ttagagttaa aggatgccca tgctacagag gagtctggag
     7321 acagcagggc tcactccagc tacctgaaga ccaagaaggg ccagtctact tcccgccata
     7381 aaaaaacaat ggtcaagaaa gtggggcctg actcagactg accggtcatc atcaccatca
     7441 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     7501 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     7561 ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     7621 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     7681 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga taccgtcgac
     7741 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7801 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct agggtgccta
     7861 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7921 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7981 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     8041 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     8101 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     8161 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     8221 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     8281 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8341 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8401 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8461 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8521 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8581 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8641 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8701 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8761 agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact cacgttaagg
     8821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     9001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     9061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     9121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     9181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     9241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9541 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9901 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg atcccctagg
     9961 gtctactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
    10021 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
    10081 cttgaccgac aattgcatga agaatctgct tagggttagg cgttt
//
