LOCUS       sequence-488804-       10857 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  Encodes fourth generation porcine endogenous retrovirus-derived 
            prime editing (pvPE) system with puromycin resistance gene into 
            mammalian cells for gene editing..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10857)
  AUTHORS   Liu W, Duan W, Peng Z, Liao Y, Wang X, Liu R, Jing Q, Jiang H, Fan 
            Y, Ge L, Huang L, Xing Y
  TITLE     Highly efficient prime editors for mammalian genome editing based on
            porcine retrovirus reverse transcriptase.
  JOURNAL   Trends Biotechnol. 2025 Aug 30:S0167-7799(25)00314-2. doi: 
            10.1016/j.tibtech.2025.07.029.
  PUBMED    40885667
REFERENCE   2  (bases 1 to 10857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Trends 
            Biotechnol. 2025 Aug 30:S0167-7799(25)00314-2. doi: 
            10.1016/j.tibtech.2025.07.029."
COMMENT     Sequence Label: pvPE-V4-Puro
FEATURES             Location/Qualifiers
     source          1..10857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     627..646
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        627..645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             695..715
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(716..3229,3230..3232,3233..4816)
                     /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             716..4816
                     /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             7394..7414
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7424..7450
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7460..7516
                     /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             7517..8116
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7517..7536)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8013..8033
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             8122..8139
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8162..8179)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8168..8392
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8463..8479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8463..8479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8476..8498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8487..8503
                     /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(8511..8517,8518..8535,8536..8541))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8556..8577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8694..8711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8865..9453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8945..8964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9624..10415,10416..10484))
                     /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     10247..10266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10485..10589)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10668..10687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat
      661 gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtcgacaa
      721 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
      781 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
      841 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
      901 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
      961 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     1021 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     1081 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     1141 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     1201 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     1261 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     1321 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     1381 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     1441 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     1501 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     1561 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     1621 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     1681 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     1741 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     1801 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     1861 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     1921 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     1981 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     2041 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     2101 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     2161 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     2221 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     2281 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     2341 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     2401 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     2461 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     2521 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     2581 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     2641 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     2701 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     2761 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     2821 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     2881 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     2941 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     3001 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     3061 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     3121 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     3181 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacg ctatcgtgcc
     3241 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     3301 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     3361 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     3421 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     3481 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     3541 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     3601 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     3661 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     3721 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     3781 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     3841 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     3901 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     3961 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     4021 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     4081 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     4141 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     4201 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     4261 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     4321 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     4381 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     4441 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     4501 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     4561 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     4621 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     4681 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     4741 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     4801 tcagctggga ggtgactctg gaggatctag cggaggatcc tctggcagcg agacaccagg
     4861 aacaagcgag tcagcaacac cagagagcag tggcggcagc agcggcggca gcagcgattt
     4921 taggaaaatt aggtcaggcc ctagacagtt agggaacctg ggcaatagga ccccactcga
     4981 caaggaccaa tgtgcatatt gtaaagaaaa aggacactgg gcaagggact gccccaagaa
     5041 gggaaacaaa ggactgaagg tcttagctct ggaagaagat aaatccggta gcgaaacacc
     5101 ggggacttca gaatcggcca ccccggagtc taccctgcag ctggatgatg aatacagact
     5161 atacagtcct ctggtgaagc cagaccagaa tatccagttc tggctcgagc agttccctca
     5221 ggcctgggct gaaaccgccg gagtgggcct ggccaaacag gtgccccctc aagtcattca
     5281 gctcaaggcc tctgccactc cagtgtctgt gcggcagaga ccgctgtcta aggaagctag
     5341 agaaggcatc cggccacacg tacaaagact gatacagcag ggcatcctgg tgcctgttca
     5401 gagcccctgg aacacccccc tgctgcctgt gagaaaacct ggaacaaatg attaccggcc
     5461 tgtgcaggac ctgagagagg tcaacaaacg ggtgcaggat atccatccta ccgtgcccaa
     5521 cccctataac ctgctgtgtg ccctgccacc acagcggagc tggtacacag ttctggatct
     5581 gaaggatgcc ttcttctgtc tgcggctcca cccaaccagc cagcctctct ttgcttttga
     5641 atggcgggac cctggcgccg gccgaacggg ccagctaacc tggacaagac tgccccaggg
     5701 ttttaaaaac tctcccacca tctttaatga agcccttcac agggacctcg ccaattttag
     5761 aatccagcac cctcaagtaa ctctgctcca gtatgctgat gacctgctac ttgccggcgc
     5821 caccaagcaa gactgtttgg aaggaaccaa agccctgctg ctggagctgt ctgacctagg
     5881 ctaccgagca agtgctaaga aggcacagat ctgccggcga gaggtcacat acctaggata
     5941 cagcctcaga ggcggccaga ggtggctcac cgaggccagg aagaggaccg tggtgcagat
     6001 cccagcccca accacggcaa agcaggtgag ggaatttctg gggaaggccg ggttctgcag
     6061 actgttcatt cctggcttcg ccaccttggc ggctcccctc tacccactta caaaggagaa
     6121 gggcgagttc tcctgggcgc cagagcatca gaaggccttt gacgccatca aaaaggctct
     6181 gctgtctgcc ccagccctgg ccttaccaga tgtcaccaag ccgttcaccc tgtatgttga
     6241 tgaaagaaaa ggggtcgcgc ggggcgtact gacccagact ctgggcccat ggagacggcc
     6301 tgtggcctac ctaagcaaga agctggaccc tgttgcctca ggctggcccg tgagactgaa
     6361 agccatagct gctgtggcca tcctggtgaa ggatgcagac aagctaactc taggtcagaa
     6421 catcaccgtc atcgcccccc acgctctgga aaacatcgtg cgccagcccc cagaccgctg
     6481 gatgaccaac gcccgcatga cacactatca gagcctctta ctgacagaga gggtgacgtt
     6541 tgctcctccc gccgccctca atcctgccac cctgctacct gaagaaaccg acgaacccgt
     6601 gacccacgac tgccaccagt tgctcatcga ggaaacagga gtcaggaaag acttgacgga
     6661 catccctcta tctggaggga gcagtggcgg cagcagcggc tctgaaacac ctggcaccag
     6721 cgagagcgcc acccccgagt cctctggtgg gagctcggga ggctccatgg ctgaaaatgg
     6781 tgataatgaa aagatggctg ccctggaggc caaaatctgt catcaaattg agtattattt
     6841 tggcgacttc aatttgccac gggacaagtt tctaaaggaa cagataaaac tggatgaagg
     6901 ctgggtacct ttggagataa tgataaaatt caacaggttg aaccgtctaa caacagactt
     6961 taatgtaatt gtggaagcat tgagcaaatc caaggcagaa ctcatggaaa tcagtgaaga
     7021 taaaactaaa atcagaaggt ctccaagcaa acccctacct gaagtgactg atgagtataa
     7081 aaatgatgta aaaaacagat ctgtttatat taaaggcttc ccaactgatg caactcttga
     7141 tgacataaaa gaatggttag aagataaagg tcaagtacta aatattcaga tgagaagaac
     7201 attgcataaa gcatttaagg gatcaatttt tgttgtgttt gatagcattg aatctgctaa
     7261 gaaatttgta gagacccctg gccagaagta caaagaaaca gacctgctaa tacttttcaa
     7321 ggacgattac tttgccaaaa aaaatgaatc tggcggctca aaaagaaccg ccgacggcag
     7381 cgaattcgag tctcccaaga agaagaggaa agtcggctct ggccctgccg ctaagagagt
     7441 gaagctggac ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga
     7501 ggagaaccct ggacctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     7561 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     7621 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     7681 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     7741 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     7801 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     7861 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca
     7921 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     7981 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     8041 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     8101 caagcccggt gcctaaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     8161 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     8221 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagaa aattgcatcg
     8281 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     8341 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     8401 gcggaaagaa ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca
     8461 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     8521 gccggaagca taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt
     8581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     8641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     8701 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     8761 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     8821 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     8881 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     8941 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     9001 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     9061 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     9121 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     9181 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     9241 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     9301 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     9361 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     9421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     9481 tctgacactc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     9541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     9601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     9661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     9721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     9781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     9841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     9901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     9961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    10021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    10081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    10141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    10201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    10261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    10321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    10381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    10441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    10501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    10561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
    10621 gacggatcgg gagatcgatc tcccgatccc ctagggtcta ctctcagtac aatctgctct
    10681 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    10741 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    10801 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgt
//
