LOCUS       sequence-390160-       10319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SpCas9 prime editor 2 under EF1-alpha 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10319)
  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 10319)
  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: pEF_PE2
FEATURES             Location/Qualifiers
     source          1..10319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..57
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             66..83
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(106..123)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    112..336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(407..423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(407..423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(420..442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    431..447
                     /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(455..461,462..479,480..485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    500..521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(638..655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(809..1397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(889..908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1568..2359,2360..2428))
                     /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     2191..2210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2429..2533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2612..2631)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2802..3983
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3032..3974
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3931..3951
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     3994..4013
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3994..4012
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4062..4082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4083..6596,6597..6599,6600..8183)
                     /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             4083..8183
                     /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 ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtctaa
       61 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
      121 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      181 ccactcccac tgtcctttcc taataaaatg agaaaattgc atcgcattgt ctgagtaggt
      241 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
      301 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
      361 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
      421 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      481 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      541 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      601 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      661 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      721 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      781 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      841 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      901 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      961 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1021 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1081 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1141 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1201 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1261 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1321 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga
     1441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1561 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1621 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1681 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1741 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1801 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1861 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1921 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1981 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2041 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2101 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2161 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2221 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2281 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2341 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2401 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2521 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     2581 gatctcccga tcccctaggg tctactctca gtacaatctg ctctgatgcc gcatagttaa
     2641 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     2701 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     2761 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tgctccggtg cccgtcagtg
     2821 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     2881 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     2941 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     3001 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     3061 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacgc ccctggctgc
     3121 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     3181 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     3241 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     3301 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     3361 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     3421 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     3481 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     3541 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     3601 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     3661 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     3721 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     3781 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     3841 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     3901 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     3961 ttcttccatt tcaggtgtcg tgaccgcggc cgctaatacg actcactata gggagagccg
     4021 ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag
     4081 tcgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     4141 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     4201 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     4261 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     4321 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     4381 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     4441 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     4501 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     4561 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     4621 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     4681 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     4741 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     4801 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     4861 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     4921 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     4981 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     5041 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     5101 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     5161 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     5221 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     5281 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     5341 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     5401 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     5461 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     5521 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     5581 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     5641 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     5701 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     5761 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     5821 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     5881 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     5941 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     6001 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     6061 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     6121 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     6181 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     6241 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     6301 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     6361 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     6421 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     6481 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     6541 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggacgcta
     6601 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     6661 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     6721 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     6781 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     6841 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     6901 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     6961 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     7021 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     7081 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     7141 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     7201 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     7261 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     7321 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     7381 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     7441 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     7501 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     7561 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     7621 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     7681 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     7741 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     7801 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     7861 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     7921 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     7981 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     8041 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     8101 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     8161 acctgtctca gctgggaggt gactctggag gatctagcgg aggatcctct ggcagcgaga
     8221 caccaggaac aagcgagtca gcaacaccag agagcagtgg cggcagcagc ggcggcagca
     8281 gcaccctaaa tatagaagat gagtatcggc tacatgagac ctcaaaagag ccagatgttt
     8341 ctctagggtc cacatggctg tctgattttc ctcaggcctg ggcggaaacc gggggcatgg
     8401 gactggcagt tcgccaagct cctctgatca tacctctgaa agcaacctct acccccgtgt
     8461 ccataaaaca ataccccatg tcacaagaag ccagactggg gatcaagccc cacatacaga
     8521 gactgttgga ccagggaata ctggtaccct gccagtcccc ctggaacacg cccctgctac
     8581 ccgttaagaa accagggact aatgattata ggcctgtcca ggatctgaga gaagtcaaca
     8641 agcgggtgga agacatccac cccaccgtgc ccaaccctta caacctcttg agcgggctcc
     8701 caccgtccca ccagtggtac actgtgcttg atttaaagga tgcctttttc tgcctgagac
     8761 tccaccccac cagtcagcct ctcttcgcct ttgagtggag agatccagag atgggaatct
     8821 caggacaatt gacctggacc agactcccac agggtttcaa aaacagtccc accctgttta
     8881 atgaggcact gcacagagac ctagcagact tccggatcca gcacccagac ttgatcctgc
     8941 tacagtacgt ggatgactta ctgctggccg ccacttctga gctagactgc caacaaggta
     9001 ctcgggccct gttacaaacc ctagggaacc tcgggtatcg ggcctcggcc aagaaagccc
     9061 aaatttgcca gaaacaggtc aagtatctgg ggtatcttct aaaagagggt cagagatggc
     9121 tgactgaggc cagaaaagag actgtgatgg ggcagcctac tccgaagacc cctcgacaac
     9181 taagggagtt cctagggaag gcaggcttct gtcgcctctt catccctggg tttgcagaaa
     9241 tggcagcccc cctgtaccct ctcaccaaac cggggactct gtttaattgg ggcccagacc
     9301 aacaaaaggc ctatcaagaa atcaagcaag ctcttctaac tgccccagcc ctggggttgc
     9361 cagatttgac taagcccttt gaactctttg tcgacgagaa gcagggctac gccaaaggtg
     9421 tcctaacgca aaaactggga ccttggcgtc ggccggtggc ctacctgtcc aaaaagctag
     9481 acccagtagc agctgggtgg cccccttgcc tacggatggt agcagccatt gccgtactga
     9541 caaaggatgc aggcaagcta accatgggac agccactagt cattctggcc ccccatgcag
     9601 tagaggcact agtcaaacaa ccccccgacc gctggctttc caacgcccgg atgactcact
     9661 atcaggcctt gcttttggac acggaccggg tccagttcgg accggtggta gccctgaacc
     9721 cggctacgct gctcccactg cctgaggaag ggctgcaaca caactgcctt gatatcctgg
     9781 ccgaagccca cggaacccga cccgacctaa cggaccagcc gctcccagac gccgaccaca
     9841 cctggtacac ggatggaagc agtctcttac aagagggaca gcgtaaggcg ggagctgcgg
     9901 tgaccaccga gaccgaggta atctgggcta aagccctgcc agccgggaca tccgctcagc
     9961 gggctgaact gatagcactc acccaggccc taaagatggc agaaggtaag aagctaaatg
    10021 tttatactga tagccgttat gcttttgcta ctgcccatat ccatggagaa atatacagaa
    10081 ggcgtgggtg gctcacatca gaaggcaaag agatcaaaaa taaagacgag atcttggccc
    10141 tactaaaagc cctctttctg cccaaaagac ttagcataat ccattgtcca ggacatcaaa
    10201 agggacacag cgccgaggct agaggcaacc ggatggctga ccaagcggcc cgaaaggcag
    10261 ccatcacaga gactccagac acctctaccc tcctcataga aaattcatca ccctctggc
//
