LOCUS       sequence-430585-       10569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prime editing in mammalian cells with co-translational GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10569)
  AUTHORS   Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, 
            Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR
  TITLE     Search-and-replace genome editing without double-strand breaks or 
            donor DNA.
  JOURNAL   Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 
            10.1038/s41586-019-1711-4.
  PUBMED    31634902
REFERENCE   2  (bases 1 to 10569)
  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: "Nature. 
            2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 
            10.1038/s41586-019-1711-4."
COMMENT     Sequence Label: pCMV-PE2-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..10569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2260..2280)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2329..2348)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2330..2348)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2390..2593)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2419..2439)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2594..2973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3145..3164
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3243..3347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3348..3416,3417..4208)
                     /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(3566..3585)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4379..4967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4868..4887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5121..5138
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5255..5276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5291..5296,5297..5314,5315..5321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5329..5345
                     /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     5334..5356
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5353..5369
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5353..5369
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5440..5664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5653..5670
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5693..5710)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5719..5739)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(5752..6462,6463..6465,6466..6468))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5752..6462,6463..6465,6466..6468))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5794..5815)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6143..6162
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(6469..6525)
                     /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(6535..6555)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(8728..10311,10312..10314,10315..10569,
                     1..2259))
                     /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(8728..10569,1..2259))
                     /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 gtgccggccc atcactttca cgagctcgtc caccaccttc actgtctgca ggatgccctt
       61 cttaatggcg gggctgccgg ccagattggc aatgtgctcg tgcaggctat cgccctggcc
      121 ggacacctgg gctttctgga tgtcctcttt aaaggtcagg ctgtcgtcgt ggatcagctg
      181 catgaagttt ctgttggcga agccgtcgga cttcaggaaa tccaggattg tcttgccgga
      241 ctgcttgtcc cggatgccgt tgatcagctt ccggctcagc ctgccccagc cggtgtatct
      301 ccgccgcttc agctgcttca tcactttgtc gtcgaacagg tgggcatagg ttttcagccg
      361 ttcctcgatc atctctctgt cctcaaacag tgtcagggtc agcacgatat cttccagaat
      421 gtcctcgttt tcctcattgt ccaggaagtc cttgtccttg ataattttca gcagatcgtg
      481 gtatgtgccc agggaggcgt tgaaccgatc ttccacgccg gagatttcca cggagtcgaa
      541 gcactcgatt ttcttgaagt agtcctcttt cagctgcttc acggtcactt tccggttggt
      601 cttgaacagc aggtccacga tggccttttt ctgctcgccg ctcaggaagg cgggctttct
      661 cattccctcg gtcacgtatt tcactttggt cagctcgtta tacacggtga agtactcgta
      721 cagcaggctg tgcttgggca gcaccttctc gttgggcagg ttcttatcga agttggtcat
      781 ccgctcgatg aagctctggg cggaagcgcc cttgtccacc acttcctcga agttccaggg
      841 ggtgatggtt tcctcgctct ttctggtcat ccaggcgaat ctgctgtttc ccctggccag
      901 agggcccacg tagtagggga tgcggaaggt caggatcttc tcgatctttt cccggttgtc
      961 cttcaggaat gggtaaaaat cttcctgccg ccgcagaatg gcgtgcagct ctcccaggtg
     1021 gatctggtgg gggatgctgc cgttgtcgaa ggtccgctgc ttccgcagca ggtcctctct
     1081 gttcagcttc acgagcagtt cctcggtgcc gtccatcttt tccaggatgg gcttgatgaa
     1141 cttgtagaac tcttcctggc tggctccgcc gtcaatgtag ccggcgtagc cgttcttgct
     1201 ctggtcgaag aaaatctctt tgtacttctc aggcagctgc tgccgcacga gagctttcag
     1261 cagggtcagg tcctggtggt gctcgtcgta tctcttgatc atagaggcgc tcaggggggc
     1321 cttggtgatc tcggtgttca ctctcaggat gtcgctcagc aggatggcgt cggacaggtt
     1381 cttggcggcc agaaacaggt cggcgtactg gtcgccgatc tgggccagca ggttgtccag
     1441 gtcgtcgtcg taggtgtcct tgctcagctg cagtttggca tcctcggcca ggtcgaagtt
     1501 gctcttgaag ttgggggtca ggcccaggct cagggcaatc aggtttccga acaggccatt
     1561 cttcttctcg ccgggcagct gggcgatcag attttccagc cgtctgctct tgctcagtct
     1621 ggcagacagg atggccttgg cgtccacgcc gctggcgttg atggggtttt cctcgaacag
     1681 ctggttgtag gtctgcacca gctggatgaa cagcttgtcc acgtcgctgt tgtcggggtt
     1741 caggtcgccc tcgatcagga agtggccccg gaacttgatc atgtgggcca gggccagata
     1801 gatcagccgc aggtcggcct tgtcggtgct gtccaccagt ttctttctca ggtggtagat
     1861 ggtggggtac ttctcgtggt aggccacctc gtccacgatg ttgccgaaga tggggtgccg
     1921 ctcgtgcttc ttatcctctt ccaccaggaa ggactcttcc agtctgtgga agaagctgtc
     1981 gtccaccttg gccatctcgt tgctgaagat ctcttgcaga tagcagatcc ggttcttccg
     2041 tctggtgtat cttcttctgg cggttctctt cagccgggtg gcctcggctg tttcgccgct
     2101 gtcgaacagc agggctccga tcaggttctt cttgatgctg tgccggtcgg tgttgcccag
     2161 caccttgaat ttcttgctgg gcaccttgta ctcgtcggtg atcacggccc agcccacaga
     2221 gttggtgccg atgtccaggc cgatgctgta cttcttgtcg actttccgct tcttctttgg
     2281 tgactcgaac tcgcttccgt cggctgtccg tttcatggtg gcggctctcc ctatagtgag
     2341 tcgtattagc ggccgcggat ctctagcgga tctgacggtt cactaaacca gctctgctta
     2401 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     2461 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     2521 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     2581 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     2641 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     2701 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     2761 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     2821 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     2881 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     2941 attactatta ataactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc
     3001 gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt
     3061 gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag
     3121 ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtagacccta
     3181 ggggatcggg agatcgatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     3241 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3301 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3361 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3421 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3481 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3541 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3601 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3661 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3721 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3781 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3841 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3901 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3961 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4021 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4081 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4141 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4201 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4261 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4321 aacgtgagtt ttcgttccac tgagtgtcag accccgtaga aaagatcaaa ggatcttctt
     4381 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4441 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4501 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4561 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4621 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4681 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4741 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4801 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4861 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4921 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4981 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5041 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5101 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5161 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     5221 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     5281 caccctaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     5341 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     5401 acggtatcga gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc
     5461 cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
     5521 ccagaataga atgacaccta ctcagacaat gcgatgcaat tttctcattt tattaggaaa
     5581 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     5641 atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaact caatggtgat
     5701 ggtgatgatg accggttaga ctttcctctt cttcttggga gaaccaccag acttgtacag
     5761 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     5821 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
     5881 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     5941 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     6001 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     6061 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     6121 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     6181 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     6241 ggggtagcgg ctgaagcact gcactccata ggtcagggtg gtcacgaggg tgggccaggg
     6301 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     6361 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     6421 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatag gtccagggtt
     6481 ctcctccacg tctccagcct gcttcagcag gctgaagtta gtagctccgc ttccgacttt
     6541 cctcttcttc ttgggctcga attcgctgcc gtcggcggtt ctttttgagc cgccagaggg
     6601 tgatgaattt tctatgagga gggtagaggt gtctggagtc tctgtgatgg ctgcctttcg
     6661 ggccgcttgg tcagccatcc ggttgcctct agcctcggcg ctgtgtccct tttgatgtcc
     6721 tggacaatgg attatgctaa gtcttttggg cagaaagagg gcttttagta gggccaagat
     6781 ctcgtcttta tttttgatct ctttgccttc tgatgtgagc cacccacgcc ttctgtatat
     6841 ttctccatgg atatgggcag tagcaaaagc ataacggcta tcagtataaa catttagctt
     6901 cttaccttct gccatcttta gggcctgggt gagtgctatc agttcagccc gctgagcgga
     6961 tgtcccggct ggcagggctt tagcccagat tacctcggtc tcggtggtca ccgcagctcc
     7021 cgccttacgc tgtccctctt gtaagagact gcttccatcc gtgtaccagg tgtggtcggc
     7081 gtctgggagc ggctggtccg ttaggtcggg tcgggttccg tgggcttcgg ccaggatatc
     7141 aaggcagttg tgttgcagcc cttcctcagg cagtgggagc agcgtagccg ggttcagggc
     7201 taccaccggt ccgaactgga cccggtccgt gtccaaaagc aaggcctgat agtgagtcat
     7261 ccgggcgttg gaaagccagc ggtcgggggg ttgtttgact agtgcctcta ctgcatgggg
     7321 ggccagaatg actagtggct gtcccatggt tagcttgcct gcatcctttg tcagtacggc
     7381 aatggctgct accatccgta ggcaaggggg ccacccagct gctactgggt ctagcttttt
     7441 ggacaggtag gccaccggcc gacgccaagg tcccagtttt tgcgttagga cacctttggc
     7501 gtagccctgc ttctcgtcga caaagagttc aaagggctta gtcaaatctg gcaaccccag
     7561 ggctggggca gttagaagag cttgcttgat ttcttgatag gccttttgtt ggtctgggcc
     7621 ccaattaaac agagtccccg gtttggtgag agggtacagg ggggctgcca tttctgcaaa
     7681 cccagggatg aagaggcgac agaagcctgc cttccctagg aactccctta gttgtcgagg
     7741 ggtcttcgga gtaggctgcc ccatcacagt ctcttttctg gcctcagtca gccatctctg
     7801 accctctttt agaagatacc ccagatactt gacctgtttc tggcaaattt gggctttctt
     7861 ggccgaggcc cgatacccga ggttccctag ggtttgtaac agggcccgag taccttgttg
     7921 gcagtctagc tcagaagtgg cggccagcag taagtcatcc acgtactgta gcaggatcaa
     7981 gtctgggtgc tggatccgga agtctgctag gtctctgtgc agtgcctcat taaacagggt
     8041 gggactgttt ttgaaaccct gtgggagtct ggtccaggtc aattgtcctg agattcccat
     8101 ctctggatct ctccactcaa aggcgaagag aggctgactg gtggggtgga gtctcaggca
     8161 gaaaaaggca tcctttaaat caagcacagt gtaccactgg tgggacggtg ggagcccgct
     8221 caagaggttg taagggttgg gcacggtggg gtggatgtct tccacccgct tgttgacttc
     8281 tctcagatcc tggacaggcc tataatcatt agtccctggt ttcttaacgg gtagcagggg
     8341 cgtgttccag ggggactggc agggtaccag tattccctgg tccaacagtc tctgtatgtg
     8401 gggcttgatc cccagtctgg cttcttgtga catggggtat tgttttatgg acacgggggt
     8461 agaggttgct ttcagaggta tgatcagagg agcttggcga actgccagtc ccatgccccc
     8521 ggtttccgcc caggcctgag gaaaatcaga cagccatgtg gaccctagag aaacatctgg
     8581 ctcttttgag gtctcatgta gccgatactc atcttctata tttagggtgc tgctgccgcc
     8641 gctgctgccg ccactgctct ctggtgttgc tgactcgctt gttcctggtg tctcgctgcc
     8701 agaggatcct ccgctagatc ctccagagtc acctcccagc tgagacaggt cgatccgtgt
     8761 ctcgtacagg ccggtgatgc tctggtggat cagggtggcg tccagcacct ctttggtgct
     8821 ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg aaggcggcag gggctcccag
     8881 attggtcagg gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg gcttatcccg
     8941 gtgcttgttg taggcggaca gcactttgtc cagattagcg tcggccagga tcactctctt
     9001 ggagaactcg ctgatctgct cgatgatctc gtccaggtag tgcttgtgct gttccacaaa
     9061 cagctgtttc tgctcattat cctcggggga gcccttcagc ttctcatagt ggctggccag
     9121 gtacaggaag ttcacatatt tggagggcag ggccagttcg tttcccttct gcagttcgcc
     9181 ggcagaggcc agcattctct tccggccgtt ttccagctcg aacagggagt acttaggcag
     9241 cttgatgatc aggtcctttt tcacttcttt gtagcccttg gcttccagaa agtcgatggg
     9301 attcttctcg aagctgcttc tttccatgat ggtgatcccc agcagctctt tcacactctt
     9361 cagtttcttg gacttgccct tttccacttt ggccaccacc agcacagaat aggccacggt
     9421 ggggctgtcg aagccgccgt acttcttagg gtcccagtcc ttctttctgg cgatcagctt
     9481 atcgctgttc ctcttgggca ggatagactc tttgctgaag ccgcctgtct gcacctcggt
     9541 ctttttcacg atattcactt ggggcatgct cagcactttc cgcacggtgg caaaatcccg
     9601 gcccttatcc cacacgatct ccccggtttc gccgtttgtc tcgatcagag gccgcttccg
     9661 gatctcgccg ttggccaggg taatctcggt cttgaaaaag ttcatgatgt tgctgtagaa
     9721 gaagtacttg gcggtagcct tgccgatttc ctgctcgctc ttggcgatca tcttccgcac
     9781 gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt actttttgat
     9841 cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt agttgttgat
     9901 ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct tggacttcag
     9961 ggtgatcact ttcacttccc ggatcagctt gtcattctcg tcgtacttag tgttcatccg
    10021 ggagtccagg atctgtgcca cgtgctttgt gatctgccgg gtttccacca gctgtctctt
    10081 gatgaagccg gccttatcca gttcgctcag gccgcctctc tcggccttgg tcagattgtc
    10141 gaactttctc tgggtaatca gcttggcgtt cagcagctgc cgccagtagt tcttcatctt
    10201 cttcacgacc tcttcggagg gcacgttgtc gctcttgccc cggttcttgt cgcttctggt
    10261 cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga tagcgtccac
    10321 atcgtagtcg gacagccggt tgatgtccag ttcctggtcc acgtacatat cccgcccatt
    10381 ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca cggggtgttc
    10441 tttcaggatc tggctgccca gctctttgat gccctcttcg atccgcttca ttctctcgcg
    10501 gctgttcttc tgtcccttct gggtggtctg gttctctctg gccatttcga tcacgatgtt
    10561 ctcgggctt
//
