LOCUS       sequence-450755-       10707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-Intron-Gag-SpyCas9-NLS. Expresses Gag-SpyCas9-NLS for VLP 
            production..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10707)
  AUTHORS   Ramadoss GN, Namaganda SJ, Kumar MM, Hamilton JR, Sharma R, Chow KG,
            Workley LA, Macklin BL, Sun M, Ha AS, Liu JC, Fellmann C, Watry HL, 
            Dierks PH, Bose RS, Jin J, Perez BS, Sandoval Espinoza CR, Matia MP,
            Lu SH, Judge LM, Shy BR, Nussenzweig A, Adamson B, Murthy N, Doudna 
            JA, Kampmann M, Conklin BR
  TITLE     Characterizing and controlling CRISPR repair outcomes in nondividing
            human cells.
  JOURNAL   Nat Commun. 2025 Nov 17;16(1):9883. doi: 10.1038/s41467-025-66058-3.
  PUBMED    41249169
REFERENCE   2  (bases 1 to 10707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-11-17"; volume: "16(1):9883. doi"; pages: " 
            10.1038/s41467-025-66058-3"
COMMENT     Sequence Label: pCF140_Gag-SpyCas9-NLS
FEATURES             Location/Qualifiers
     source          1..10707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             3100..3123
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3130..3150
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3175..7275
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3175..7275
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7276..7323
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7412..7431)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7477..7532
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7531..7550)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7890..7906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7890..7906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7903..7925)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7914..7930
                     /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(7938..7944,7945..7962,7963..7968))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7983..8004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8121..8138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8292..8880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8372..8391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9051..9842,9843..9911))
                     /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     9674..9693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9912..10016)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10042..10497)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10179..10200)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10391..10410
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10624..10646
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10638..10655
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10639..10655
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10665..10684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10665..10683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      721 gctgttttga cctccataga agacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattcct cgagggccgg atccgaattc
     1441 ctgtatttgt ctgaaaacat gggccaggct gttaccaccc ccttaagttt gactttagac
     1501 cactggaagg atgtcgaacg gacagcccac aacctgtcgg tagaggttag aaaaaggcgc
     1561 tgggttacat tctgctctgc agaatggcca accttcaacg tcggatggcc acgagacggc
     1621 acttttaacc cagacattat tacacaggtt aagatcaagg tcttctcacc tggcccacat
     1681 ggacatccgg atcaggtccc ctacatcgtg acctgggaag ctatagcagt agacccccct
     1741 ccctgggtca gacccttcgt gcaccctaaa cctcccctct ctcttccccc ttcagccccc
     1801 tctctcccac ctgaaccccc actctcgacc ccgccccagt cctccctcta tccggctctc
     1861 acttctcctt taaacaccaa acctaggcct caagtccttc ctgatagcgg aggaccactc
     1921 attgatctac tcacggagga ccctccgcct taccgggacc cagggccacc ctctcctgac
     1981 gggaacggcg atagcggaga agtggcccct acagaaggag cccctgaccc ttccccaatg
     2041 gtatcccgcc tgcggggaag aaaagaaccc cccgtggcgg attctactac ctctcaggcg
     2101 ttcccccttc gcctgggagg gaatggacag tatcaatact ggccattttc ctcctctgac
     2161 ctctataact ggaaaaataa caacccctct ttctccgagg acccagctaa attgacagct
     2221 ttgatcgagt ccgttctcct tactcatcag cccacttggg atgactgcca acagctatta
     2281 gggaccctgc tgacgggaga agaaaaacag cgagtgctcc tagaggcccg aaaggcggtt
     2341 cgaggggagg acggacgccc aactcagctg cccaatgaca ttaatgatgc ttttcccttg
     2401 gaacgtcccg actgggacta caacacccaa cgaggtagga accacctagt ccactatcgc
     2461 cagttgctcc tagcgggtct ccaaaacgcg ggcagaagcc ccaccaattt ggccaaggta
     2521 aaagggataa cccagggacc taatgagtct ccctcagcct ttttagagag actcaaggag
     2581 gcctatcgca gatacactcc ttatgaccct gaggacccag ggcaagaaac caatgtggcc
     2641 atgtcattca tctggcagtc cgccccggat atcgggcgaa agttagagcg gttagaagat
     2701 ttgaagagta agaccttagg agacttagtg agggaagctg aaaagatctt taataaacga
     2761 gaaaccccgg aagaaagaga ggaacgtatt aggagagaaa cagaggaaaa ggaagaacgc
     2821 cgtagggcag aggatgtgca gagagagaag gagagggacc gcagaagaca tagagaaatg
     2881 agtaagttgc tggctactgt cgttagcggg cagagacagg atagacaggg aggagagcga
     2941 aggaggcccc aactcgacca cgaccagtgt gcctactgca aagaaaaggg acattgggct
     3001 agagattgcc ccaagaagcc aagaggaccc cggggaccac gaccccaggc ctccctcctg
     3061 acgcgtagtt ccctgtatcc agccctcaca cctaccggtg attacaaaga cgatgacgat
     3121 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
     3181 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     3241 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     3301 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     3361 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     3421 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     3481 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     3541 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     3601 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     3661 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     3721 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     3781 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     3841 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     3901 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     3961 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     4021 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     4081 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     4141 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     4201 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     4261 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     4321 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     4381 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     4441 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     4501 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     4561 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     4621 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     4681 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     4741 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     4801 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     4861 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     4921 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     4981 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     5041 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     5101 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     5161 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     5221 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     5281 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     5341 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     5401 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     5461 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     5521 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     5581 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     5641 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     5701 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     5761 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     5821 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     5881 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     5941 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     6001 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     6061 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     6121 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     6181 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     6241 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     6301 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     6361 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     6421 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     6481 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     6541 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     6601 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     6661 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     6721 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     6781 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     6841 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     6901 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     6961 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     7021 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     7081 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     7141 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     7201 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     7261 cagctgggag gcgacaagcg tcctgctgct actaagaaag ctggtcaagc taagaaaaag
     7321 aaatgaggta cctgcatcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     7381 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     7441 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     7501 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     7561 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     7621 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     7681 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     7741 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     7801 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     7861 tcgacggatc ggccgcaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     7921 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7981 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     8041 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     8101 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     8161 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8221 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8281 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8341 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8401 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8461 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8521 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8581 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8641 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8701 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     8761 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8821 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8881 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8941 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9001 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9061 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9121 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9181 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9241 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9301 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9361 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9421 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9481 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9541 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9601 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9661 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9721 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9781 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9841 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9901 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9961 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10021 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
    10081 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
    10141 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
    10201 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
    10261 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
    10321 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
    10381 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
    10441 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
    10501 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
    10561 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
    10621 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
    10681 agggcgaatt ggagctccac cgcggtg
//
