LOCUS       sequence-440213-       10423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PE2 with puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10423)
  AUTHORS   Koeppel J, Weller J, Peets EM, Pallaseni A, Kuzmin I, Raudvere U, 
            Peterson H, Liberante FG, Parts L
  TITLE     Prediction of prime editing insertion efficiencies using sequence 
            features and DNA repair determinants.
  JOURNAL   Nat Biotechnol. 2023 Feb 16. doi: 10.1038/s41587-023-01678-y.
  PUBMED    36797492
REFERENCE   2  (bases 1 to 10423)
  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 
            Biotechnol. 2023 Feb 16. doi: 10.1038/s41587-023-01678-y."
COMMENT     Sequence Label: pCMV-PE2-P2A-PuroR
FEATURES             Location/Qualifiers
     source          1..10423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3601..3621)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3670..3689)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3671..3689)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3731..3934)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3760..3780)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3935..4314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4486..4505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4584..4688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4689..4757,4758..5549)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4907..4926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5720..6308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6209..6228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6462..6479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6596..6617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6632..6637,6638..6655,6656..6662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6670..6686
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6675..6697
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6694..6710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6694..6710
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6781..7005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6994..7011
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(7034..7051)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     7644..7663
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(7664..7720)
                     /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             complement(7730..7750)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(9923..10423,1..1083,1084..1086,1087..3600))
                     /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             complement(join(9923..10423,1..3600))
                     /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 gtacttaggc agcttgatga tcaggtcctt tttcacttct ttgtagccct tggcttccag
       61 aaagtcgatg ggattcttct cgaagctgct tctttccatg atggtgatcc ccagcagctc
      121 tttcacactc ttcagtttct tggacttgcc cttttccact ttggccacca ccagcacaga
      181 ataggccacg gtggggctgt cgaagccgcc gtacttctta gggtcccagt ccttctttct
      241 ggcgatcagc ttatcgctgt tcctcttggg caggatagac tctttgctga agccgcctgt
      301 ctgcacctcg gtctttttca cgatattcac ttggggcatg ctcagcactt tccgcacggt
      361 ggcaaaatcc cggcccttat cccacacgat ctccccggtt tcgccgtttg tctcgatcag
      421 aggccgcttc cggatctcgc cgttggccag ggtaatctcg gtcttgaaaa agttcatgat
      481 gttgctgtag aagaagtact tggcggtagc cttgccgatt tcctgctcgc tcttggcgat
      541 catcttccgc acgtcgtaca ccttgtagtc gccgtacacg aactcgcttt ccagcttagg
      601 gtactttttg atcagggcgg ttcccacgac ggcgttcagg taggcgtcgt gggcgtggtg
      661 gtagttgttg atctcgcgca ctttgtaaaa ctggaaatcc ttccggaaat cggacaccag
      721 cttggacttc agggtgatca ctttcacttc ccggatcagc ttgtcattct cgtcgtactt
      781 agtgttcatc cgggagtcca ggatctgtgc cacgtgcttt gtgatctgcc gggtttccac
      841 cagctgtctc ttgatgaagc cggccttatc cagttcgctc aggccgcctc tctcggcctt
      901 ggtcagattg tcgaactttc tctgggtaat cagcttggcg ttcagcagct gccgccagta
      961 gttcttcatc ttcttcacga cctcttcgga gggcacgttg tcgctcttgc cccggttctt
     1021 gtcgcttctg gtcagcacct tgttgtcgat ggagtcgtcc ttcagaaagc tctgaggcac
     1081 gatagcgtcc acatcgtagt cggacagccg gttgatgtcc agttcctggt ccacgtacat
     1141 atcccgccca ttctgcaggt agtacaggta cagcttctcg ttctgcagct gggtgttttc
     1201 cacggggtgt tctttcagga tctggctgcc cagctctttg atgccctctt cgatccgctt
     1261 cattctctcg cggctgttct tctgtccctt ctgggtggtc tggttctctc tggccatttc
     1321 gatcacgatg ttctcgggct tgtgccggcc catcactttc acgagctcgt ccaccacctt
     1381 cactgtctgc aggatgccct tcttaatggc ggggctgccg gccagattgg caatgtgctc
     1441 gtgcaggcta tcgccctggc cggacacctg ggctttctgg atgtcctctt taaaggtcag
     1501 gctgtcgtcg tggatcagct gcatgaagtt tctgttggcg aagccgtcgg acttcaggaa
     1561 atccaggatt gtcttgccgg actgcttgtc ccggatgccg ttgatcagct tccggctcag
     1621 cctgccccag ccggtgtatc tccgccgctt cagctgcttc atcactttgt cgtcgaacag
     1681 gtgggcatag gttttcagcc gttcctcgat catctctctg tcctcaaaca gtgtcagggt
     1741 cagcacgata tcttccagaa tgtcctcgtt ttcctcattg tccaggaagt ccttgtcctt
     1801 gataattttc agcagatcgt ggtatgtgcc cagggaggcg ttgaaccgat cttccacgcc
     1861 ggagatttcc acggagtcga agcactcgat tttcttgaag tagtcctctt tcagctgctt
     1921 cacggtcact ttccggttgg tcttgaacag caggtccacg atggcctttt tctgctcgcc
     1981 gctcaggaag gcgggctttc tcattccctc ggtcacgtat ttcactttgg tcagctcgtt
     2041 atacacggtg aagtactcgt acagcaggct gtgcttgggc agcaccttct cgttgggcag
     2101 gttcttatcg aagttggtca tccgctcgat gaagctctgg gcggaagcgc ccttgtccac
     2161 cacttcctcg aagttccagg gggtgatggt ttcctcgctc tttctggtca tccaggcgaa
     2221 tctgctgttt cccctggcca gagggcccac gtagtagggg atgcggaagg tcaggatctt
     2281 ctcgatcttt tcccggttgt ccttcaggaa tgggtaaaaa tcttcctgcc gccgcagaat
     2341 ggcgtgcagc tctcccaggt ggatctggtg ggggatgctg ccgttgtcga aggtccgctg
     2401 cttccgcagc aggtcctctc tgttcagctt cacgagcagt tcctcggtgc cgtccatctt
     2461 ttccaggatg ggcttgatga acttgtagaa ctcttcctgg ctggctccgc cgtcaatgta
     2521 gccggcgtag ccgttcttgc tctggtcgaa gaaaatctct ttgtacttct caggcagctg
     2581 ctgccgcacg agagctttca gcagggtcag gtcctggtgg tgctcgtcgt atctcttgat
     2641 catagaggcg ctcagggggg ccttggtgat ctcggtgttc actctcagga tgtcgctcag
     2701 caggatggcg tcggacaggt tcttggcggc cagaaacagg tcggcgtact ggtcgccgat
     2761 ctgggccagc aggttgtcca ggtcgtcgtc gtaggtgtcc ttgctcagct gcagtttggc
     2821 atcctcggcc aggtcgaagt tgctcttgaa gttgggggtc aggcccaggc tcagggcaat
     2881 caggtttccg aacaggccat tcttcttctc gccgggcagc tgggcgatca gattttccag
     2941 ccgtctgctc ttgctcagtc tggcagacag gatggccttg gcgtccacgc cgctggcgtt
     3001 gatggggttt tcctcgaaca gctggttgta ggtctgcacc agctggatga acagcttgtc
     3061 cacgtcgctg ttgtcggggt tcaggtcgcc ctcgatcagg aagtggcccc ggaacttgat
     3121 catgtgggcc agggccagat agatcagccg caggtcggcc ttgtcggtgc tgtccaccag
     3181 tttctttctc aggtggtaga tggtggggta cttctcgtgg taggccacct cgtccacgat
     3241 gttgccgaag atggggtgcc gctcgtgctt cttatcctct tccaccagga aggactcttc
     3301 cagtctgtgg aagaagctgt cgtccacctt ggccatctcg ttgctgaaga tctcttgcag
     3361 atagcagatc cggttcttcc gtctggtgta tcttcttctg gcggttctct tcagccgggt
     3421 ggcctcggct gtttcgccgc tgtcgaacag cagggctccg atcaggttct tcttgatgct
     3481 gtgccggtcg gtgttgccca gcaccttgaa tttcttgctg ggcaccttgt actcgtcggt
     3541 gatcacggcc cagcccacag agttggtgcc gatgtccagg ccgatgctgt acttcttgtc
     3601 gactttccgc ttcttctttg gtgactcgaa ctcgcttccg tcggctgtcc gtttcatggt
     3661 ggcggctctc cctatagtga gtcgtattag cggccgcgga tctctagcgg atctgacggt
     3721 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     3781 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     3841 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     3901 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     3961 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4021 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4081 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     4141 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     4201 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     4261 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcaacgcg
     4321 tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca
     4381 tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag
     4441 cgacctccaa cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga
     4501 ttgtactgag agtagaccct aggggatcgg gagatcgatc tcccgatccg tcgacgtcag
     4561 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4621 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4681 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4741 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4801 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4861 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4921 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4981 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5041 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5101 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5161 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5221 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5281 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5341 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5401 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5461 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5521 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5581 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5641 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagtgtca gaccccgtag
     5701 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5761 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5821 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5881 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5941 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6001 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6061 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6181 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6241 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6301 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6361 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6421 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6481 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6541 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6601 tgagttagct cactcattag gcaccctagg ctttacactt tatgcttccg gctcgtatgt
     6661 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6721 ccaagctcta gctagaggtc gacggtatcg agccccagct ggttctttcc gcctcagaag
     6781 ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc
     6841 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
     6901 ttttctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
     6961 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc
     7021 gggtttaaac tcaatggtga tggtgatgat gaccggtcgt ctcttaggca ccgggcttgc
     7081 gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc
     7141 cgagccgctc gtagaagggg aggtttctgg gagcggaggt ttcaaggaag gctgggaccc
     7201 cggccctctc tgccgcttcc actccaggga gcactacagc gctgcccaga cccttgcctt
     7261 ggtggtcggg cgatacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg
     7321 gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca
     7381 ttctaggccc gatttctgcg aacacggctc cagcttcaac gctctcgggt gtggtccaga
     7441 cagctaccgc ggcgccatcg tctgccaccc acaccttgcc gatgtcgagc ccgacgcgcg
     7501 tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg tctgtctgga tcgacagtgt
     7561 gacgggttgc ggggtagtca gcgaacgcgg cggccagggt ccgcacggcc ctaggaacgt
     7621 catctcgtgt agcgaggcgc accgtgggct tgtactcggt cataggtcca gggttctcct
     7681 ccacgtctcc agcctgcttc agcaggctga agttagtagc tccgcttccg actttcctct
     7741 tcttcttggg ctcgaattcg ctgccgtcgg cggttctttt tgagccgcca gagggtgatg
     7801 aattttctat gaggagggta gaggtgtctg gagtctctgt gatggctgcc tttcgggccg
     7861 cttggtcagc catccggttg cctctagcct cggcgctgtg tcccttttga tgtcctggac
     7921 aatggattat gctaagtctt ttgggcagaa agagggcttt tagtagggcc aagatctcgt
     7981 ctttattttt gatctctttg ccttctgatg tgagccaccc acgccttctg tatatttctc
     8041 catggatatg ggcagtagca aaagcataac ggctatcagt ataaacattt agcttcttac
     8101 cttctgccat ctttagggcc tgggtgagtg ctatcagttc agcccgctga gcggatgtcc
     8161 cggctggcag ggctttagcc cagattacct cggtctcggt ggtcaccgca gctcccgcct
     8221 tacgctgtcc ctcttgtaag agactgcttc catccgtgta ccaggtgtgg tcggcgtctg
     8281 ggagcggctg gtccgttagg tcgggtcggg ttccgtgggc ttcggccagg atatcaaggc
     8341 agttgtgttg cagcccttcc tcaggcagtg ggagcagcgt agccgggttc agggctacca
     8401 ccggtccgaa ctggacccgg tccgtgtcca aaagcaaggc ctgatagtga gtcatccggg
     8461 cgttggaaag ccagcggtcg gggggttgtt tgactagtgc ctctactgca tggggggcca
     8521 gaatgactag tggctgtccc atggttagct tgcctgcatc ctttgtcagt acggcaatgg
     8581 ctgctaccat ccgtaggcaa gggggccacc cagctgctac tgggtctagc tttttggaca
     8641 ggtaggccac cggccgacgc caaggtccca gtttttgcgt taggacacct ttggcgtagc
     8701 cctgcttctc gtcgacaaag agttcaaagg gcttagtcaa atctggcaac cccagggctg
     8761 gggcagttag aagagcttgc ttgatttctt gataggcctt ttgttggtct gggccccaat
     8821 taaacagagt ccccggtttg gtgagagggt acaggggggc tgccatttct gcaaacccag
     8881 ggatgaagag gcgacagaag cctgccttcc ctaggaactc ccttagttgt cgaggggtct
     8941 tcggagtagg ctgccccatc acagtctctt ttctggcctc agtcagccat ctctgaccct
     9001 cttttagaag ataccccaga tacttgacct gtttctggca aatttgggct ttcttggccg
     9061 aggcccgata cccgaggttc cctagggttt gtaacagggc ccgagtacct tgttggcagt
     9121 ctagctcaga agtggcggcc agcagtaagt catccacgta ctgtagcagg atcaagtctg
     9181 ggtgctggat ccggaagtct gctaggtctc tgtgcagtgc ctcattaaac agggtgggac
     9241 tgtttttgaa accctgtggg agtctggtcc aggtcaattg tcctgagatt cccatctctg
     9301 gatctctcca ctcaaaggcg aagagaggct gactggtggg gtggagtctc aggcagaaaa
     9361 aggcatcctt taaatcaagc acagtgtacc actggtggga cggtgggagc ccgctcaaga
     9421 ggttgtaagg gttgggcacg gtggggtgga tgtcttccac ccgcttgttg acttctctca
     9481 gatcctggac aggcctataa tcattagtcc ctggtttctt aacgggtagc aggggcgtgt
     9541 tccaggggga ctggcagggt accagtattc cctggtccaa cagtctctgt atgtggggct
     9601 tgatccccag tctggcttct tgtgacatgg ggtattgttt tatggacacg ggggtagagg
     9661 ttgctttcag aggtatgatc agaggagctt ggcgaactgc cagtcccatg cccccggttt
     9721 ccgcccaggc ctgaggaaaa tcagacagcc atgtggaccc tagagaaaca tctggctctt
     9781 ttgaggtctc atgtagccga tactcatctt ctatatttag ggtgctgctg ccgccgctgc
     9841 tgccgccact gctctctggt gttgctgact cgcttgttcc tggtgtctcg ctgccagagg
     9901 atcctccgct agatcctcca gagtcacctc ccagctgaga caggtcgatc cgtgtctcgt
     9961 acaggccggt gatgctctgg tggatcaggg tggcgtccag cacctctttg gtgctggtgt
    10021 acctcttccg gtcgatggtg gtgtcaaagt acttgaaggc ggcaggggct cccagattgg
    10081 tcagggtaaa caggtggatg atattctcgg cctgctctct gatgggctta tcccggtgct
    10141 tgttgtaggc ggacagcact ttgtccagat tagcgtcggc caggatcact ctcttggaga
    10201 actcgctgat ctgctcgatg atctcgtcca ggtagtgctt gtgctgttcc acaaacagct
    10261 gtttctgctc attatcctcg ggggagccct tcagcttctc atagtggctg gccaggtaca
    10321 ggaagttcac atatttggag ggcagggcca gttcgtttcc cttctgcagt tcgccggcag
    10381 aggccagcat tctcttccgg ccgttttcca gctcgaacag gga
//
