LOCUS       sequence-377009-       11006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PE2 in mammalian cells with co-expression of tagRFP 
            and Zeocin resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11006)
  AUTHORS   Mathis N, Allam A, Kissling L, Marquart KF, Schmidheini L, Solari C,
            Balazs Z, Krauthammer M, Schwank G
  TITLE     Predicting prime editing efficiency and product purity by deep 
            learning.
  JOURNAL   Nat Biotechnol. 2023 Jan 16. doi: 10.1038/s41587-022-01613-7.
  PUBMED    36646933
REFERENCE   2  (bases 1 to 11006)
  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 Jan 16. doi: 10.1038/s41587-022-01613-7."
COMMENT     Sequence Label: pCMV-PE2-tagRFP-BleoR
FEATURES             Location/Qualifiers
     source          1..11006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4972..4992
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5002..5058
                     /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             5059..5769
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             5779..5835
                     /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             5836..6210
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             6225..6245
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6254..6271
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6294..6311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6300..6524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6595..6611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6595..6611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6608..6630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6619..6635
                     /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(6643..6649,6650..6667,6668..6673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6688..6709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6826..6843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6997..7585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7077..7096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7756..8547,8548..8616))
                     /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     8379..8398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8617..8721)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8800..8819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8991..9370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9371..9574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9525..9545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9616..9635
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9616..9634
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             9684..9704
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(9705..11006,1..1212,1213..1215,1216..2799)
                     /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             join(9705..11006,1..2799)
                     /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 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
       61 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
      121 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
      181 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
      241 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
      301 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
      361 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
      421 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
      481 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
      541 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
      601 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
      661 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
      721 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
      781 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
      841 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
      901 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
      961 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     1021 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     1081 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     1141 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     1201 tacgatgtgg acgctatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     1261 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     1321 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     1381 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     1441 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     1501 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     1561 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     1621 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     1681 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     1741 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     1801 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     1861 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     1921 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     1981 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     2041 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     2101 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     2161 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     2221 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     2281 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     2341 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     2401 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     2461 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     2521 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     2581 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     2641 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     2701 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     2761 tacgagacac ggatcgacct gtctcagctg ggaggtgact ctggaggatc tagcggagga
     2821 tcctctggca gcgagacacc aggaacaagc gagtcagcaa caccagagag cagtggcggc
     2881 agcagcggcg gcagcagcac cctaaatata gaagatgagt atcggctaca tgagacctca
     2941 aaagagccag atgtttctct agggtccaca tggctgtctg attttcctca ggcctgggcg
     3001 gaaaccgggg gcatgggact ggcagttcgc caagctcctc tgatcatacc tctgaaagca
     3061 acctctaccc ccgtgtccat aaaacaatac cccatgtcac aagaagccag actggggatc
     3121 aagccccaca tacagagact gttggaccag ggaatactgg taccctgcca gtccccctgg
     3181 aacacgcccc tgctacccgt taagaaacca gggactaatg attataggcc tgtccaggat
     3241 ctgagagaag tcaacaagcg ggtggaagac atccacccca ccgtgcccaa cccttacaac
     3301 ctcttgagcg ggctcccacc gtcccaccag tggtacactg tgcttgattt aaaggatgcc
     3361 tttttctgcc tgagactcca ccccaccagt cagcctctct tcgcctttga gtggagagat
     3421 ccagagatgg gaatctcagg acaattgacc tggaccagac tcccacaggg tttcaaaaac
     3481 agtcccaccc tgtttaatga ggcactgcac agagacctag cagacttccg gatccagcac
     3541 ccagacttga tcctgctaca gtacgtggat gacttactgc tggccgccac ttctgagcta
     3601 gactgccaac aaggtactcg ggccctgtta caaaccctag ggaacctcgg gtatcgggcc
     3661 tcggccaaga aagcccaaat ttgccagaaa caggtcaagt atctggggta tcttctaaaa
     3721 gagggtcaga gatggctgac tgaggccaga aaagagactg tgatggggca gcctactccg
     3781 aagacccctc gacaactaag ggagttccta gggaaggcag gcttctgtcg cctcttcatc
     3841 cctgggtttg cagaaatggc agcccccctg taccctctca ccaaaccggg gactctgttt
     3901 aattggggcc cagaccaaca aaaggcctat caagaaatca agcaagctct tctaactgcc
     3961 ccagccctgg ggttgccaga tttgactaag ccctttgaac tctttgtcga cgagaagcag
     4021 ggctacgcca aaggtgtcct aacgcaaaaa ctgggacctt ggcgtcggcc ggtggcctac
     4081 ctgtccaaaa agctagaccc agtagcagct gggtggcccc cttgcctacg gatggtagca
     4141 gccattgccg tactgacaaa ggatgcaggc aagctaacca tgggacagcc actagtcatt
     4201 ctggcccccc atgcagtaga ggcactagtc aaacaacccc ccgaccgctg gctttccaac
     4261 gcccggatga ctcactatca ggccttgctt ttggacacgg accgggtcca gttcggaccg
     4321 gtggtagccc tgaacccggc tacgctgctc ccactgcctg aggaagggct gcaacacaac
     4381 tgccttgata tcctggccga agcccacgga acccgacccg acctaacgga ccagccgctc
     4441 ccagacgccg accacacctg gtacacggat ggaagcagtc tcttacaaga gggacagcgt
     4501 aaggcgggag ctgcggtgac caccgagacc gaggtaatct gggctaaagc cctgccagcc
     4561 gggacatccg ctcagcgggc tgaactgata gcactcaccc aggccctaaa gatggcagaa
     4621 ggtaagaagc taaatgttta tactgatagc cgttatgctt ttgctactgc ccatatccat
     4681 ggagaaatat acagaaggcg tgggtggctc acatcagaag gcaaagagat caaaaataaa
     4741 gacgagatct tggccctact aaaagccctc tttctgccca aaagacttag cataatccat
     4801 tgtccaggac atcaaaaggg acacagcgcc gaggctagag gcaaccggat ggctgaccaa
     4861 gcggcccgaa aggcagccat cacagagact ccagacacct ctaccctcct catagaaaat
     4921 tcatcaccct ctggcggctc aaaaagaacc gccgacggca gcgaattcga gcccaagaag
     4981 aagaggaaag tcggaagcgg cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc
     5041 gaagagaatc ctggaccgat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac
     5101 atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc
     5161 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
     5221 ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc
     5281 aaccacaccc agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg
     5341 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
     5401 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc
     5461 cctgtgatgc agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct
     5521 gacggcggcc tggaaggcag aagcgacatg gccctgaagc tcgtgggcgg gggccacctg
     5581 atctgcaact tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc
     5641 ggcgtctact atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac
     5701 gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac
     5761 aaacttaatg gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa
     5821 gagaatcctg gaccgatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     5881 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     5941 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     6001 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     6061 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     6121 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     6181 gtgcacttcg tggccgagga gcaggactga attaagaatt cgagcccaag aagaagagga
     6241 aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     6301 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6361 ggaaggtgcc actcccactg tcctttccta ataaaatgag aaaattgcat cgcattgtct
     6421 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6481 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6541 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6661 cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacac
     7621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8761 gggagatcga tctcccgatc ccctagggtc tactctcagt acaatctgct ctgatgccgc
     8821 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     8881 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     8941 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     9001 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     9061 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     9121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     9181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     9241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     9301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     9361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     9421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     9481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     9541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     9601 gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg aaacggacag
     9661 ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag aagtacagca
     9721 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     9781 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     9841 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     9901 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     9961 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
    10021 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
    10081 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
    10141 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
    10201 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
    10261 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
    10321 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
    10381 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
    10441 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
    10501 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
    10561 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
    10621 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
    10681 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
    10741 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
    10801 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
    10861 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
    10921 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
    10981 aagattttta cccattcctg aaggac
//
