pDY1330 STITCHR pCMV-nCas9-XTEN-v170_R2Tocc

LOCUS       pDY1330 STITCHR pCMV-nCas9-XTEN-v170_R2Tocc     11373 bp    DNA     circular SYN 14-APR-2025
DEFINITION  STITCHR R2Tocc editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11373)
  AUTHORS   Fell CW, Villiger L, Lim J, Hiraizumi M, Tagliaferri D, Yarnall MTN,
            Lee A, Jiang K, Kayabolen A, Krajeski RN, Schmitt-Ulms C, Ramani H, 
            Yousef SM, Roberts N, Vakulskas CA, Nishimasu H, Abudayyeh OO, 
            Gootenberg JS
  TITLE     Reprogramming site-specific retrotransposon activity to new DNA 
            sites.
  JOURNAL   Nature. 2025 Apr 9. doi: 10.1038/s41586-025-08877-4.
  PUBMED    40205048
REFERENCE   2  (bases 1 to 11373)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2025 Apr 9. doi: 10.1038/s41586-025-08877-4."
COMMENT     Sequence Label: pDY1330 STITCHR pCMV-nCas9-XTEN-v170_R2Tocc
FEATURES             Location/Qualifiers
     source          1..11373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(77..96)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        268..647
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        648..851
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     802..822
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     893..912
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        893..911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             961..981
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             994..1020
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             join(1045..3558,3559..3561,3562..5145)
                     /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             1045..5145
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5167..5187
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8815..8835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8863..8889
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             8904..8921
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8944..8961)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8950..9174
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9245..9261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9245..9261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9258..9280)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9269..9285
                     /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(9293..9299,9300..9317,9318..9323))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9338..9359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9476..9493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9647..10235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9727..9746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10406..11197,11198..11266))
                     /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     11029..11048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11267..11371)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
       61 tagggtctac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
      121 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
      181 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
      241 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      301 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      361 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      421 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      481 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      541 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      601 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      661 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      721 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      781 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      841 taagcagagc tggtttagtg aaccgtcaga tccgctagag atccgcggcc gctaatacga
      901 ctcactatag ggagagccgc caccatgaaa cggacagccg acggaagcga gttcgagtca
      961 ccaaagaaga agcggaaagt ctctggggga agccccgcag cgaagcgagt gaagttggac
     1021 tcaggaggat ctagcggagg gagcgacaag aagtacagca tcggcctgga catcggcacc
     1081 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     1141 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     1201 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     1261 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     1321 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     1381 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     1441 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     1501 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     1561 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     1621 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     1681 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     1741 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     1801 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     1861 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     1921 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     1981 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     2041 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     2101 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     2161 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     2221 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     2281 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     2341 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     2401 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     2461 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     2521 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     2581 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     2641 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     2701 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     2761 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     2821 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     2881 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     2941 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     3001 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     3061 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     3121 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     3181 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     3241 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     3301 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     3361 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     3421 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     3481 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     3541 tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     3601 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     3661 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     3721 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     3781 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     3841 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     3901 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     3961 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     4021 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     4081 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     4141 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     4201 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     4261 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     4321 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     4381 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     4441 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     4501 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     4561 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     4621 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     4681 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     4741 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     4801 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     4861 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     4921 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     4981 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     5041 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     5101 ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg aggatccagc
     5161 ggatccccca agaaaaagcg caaggtatct ggaggatccg agactcccgg aacttccgaa
     5221 tctgccaccc ccgagtcctc tggaagcgag accccaggca catccgaaag cgccacgcca
     5281 gaaagcagcg gcggctcggt gaaagtgacc gtgcctgata aaaatccgcc atgcccatgt
     5341 tgttctacta gactgaacag cgtcctcgca cttatcgacc acttgaaggg tagccatggg
     5401 aagcgtaggg tgtgttttag atgtgcgaaa tgtggtcggg aaaactttaa tcaccatagt
     5461 actgtgtgtc actttgccaa gtgtaagggg ccttctgagg agaagcctcc tgtaggcgag
     5521 tggatctgtg aagtgtgcgg cagggatttc accaccaaaa taggtctcgg acagcacaaa
     5581 agactggctc acccgatggt gaggaaccag gagcggattg atgcaagcca gcctaaggag
     5641 acatcaaatc gcggcgctca caagaaatgc tggacaaagg aagaagaaga gctgcttgct
     5701 agacttgagg tgcagtttga ggggcacaag aacattaata agctcatcgc tgaacacatt
     5761 acaaccaaga ctaacaagca gatcagtgac aagcgacgcc aaatgacccg gaaggataag
     5821 ggagaaggcg gagcggcagg caagctggga cctgacactg ggaggggcaa ccactcacag
     5881 gccaaagtgg gaaataacgg tctcggaggc aatcagctgc ccggtggacc tgctgccact
     5941 aaggacaagg ctggctgcca tttagataag gaggagggga atcgaattgc gatttcacag
     6001 cagaagaaag gccgattaca gggacgctat cacaaagaaa tcaaacgacg gctcgaagag
     6061 ggcgtgatca atacgttcac caaggctttc aagcagctcc tggaatgcca ggaagtacag
     6121 cccttgataa acaagactgc acaggactgt tttgggttac tggaaagcgc atgtcacatc
     6181 cgcaccgcac tgagggggaa gaacaagaaa gaaacccaag aaaagcctac cgggggccag
     6241 tgtctgaagt ggatgaaaaa gcgcgccgtg aaaaagggca actatttgcg gtttcagcgg
     6301 ttgtttcacc ttgatagggg caagctggcg cggatcatcc ttgacgatat tgagtgtctg
     6361 tcatgtgata tcgcgccaag cgagatctac agcgtgttca aggctagatg ggagaccccc
     6421 ggacagtttg ctggtttggg caatttcaag tctaccggaa aggctgacaa taaggctttt
     6481 tccgatctga tcactgctaa ggagatcaag aagaatgtac aggagatgag caaggggagt
     6541 gctccgggtc cagatggtat cgccatagga gacattaagg gaatggaccc aggctactca
     6601 cgcactgcag agctgtttaa tctgtggtta acctctggtg agatacccga catggttaga
     6661 gggtgcagga ccgtcctgat tcctaaaagt acccagccag aacgcctcaa ggatattaac
     6721 aattggagac ccattacaat cgggtctata ctcctgagac ttttttctcg catcattacc
     6781 gctcggatga caaaggcgtg tcccttgaac cctagacaga gggggttcat cagagcagcc
     6841 ggatgttcag aaaacctgaa gctgttgcag acgattatta ggacggcgaa gtccgagcac
     6901 aggccactcg gcgtagtatt tgttgatatc gcaaaggcgt ttgataccgt gagccatcaa
     6961 catattcttc atgtgcttca gcagagaggg gtagaccccc acattatagg cttggttagt
     7021 aacatgtaca aggatatatc cacgttcgta actactaaga aagacaccca cacagacaaa
     7081 atccagatac gcgtaggtgt taagcaaggc gaccctttat ctccgttact gtttaacctg
     7141 gcaatggacc ccctcttgtg caaattagag gagagtggca acgggttcca caggggcgga
     7201 catacgatca ctgctatggc ttttgctgat gacctggtcc tccttagtga ctcttgggag
     7261 aacatggaga aaaatattga gatcttggaa gccttttgtg atctcactgg tctgaaaact
     7321 caggggcaaa agtgtcatgg attctatatc aagcctacta aagattctta tacagtgaac
     7381 aactgtgcgg cttggaccat ttacggtacc ccactgaata tgatcaatcc tggcgattcc
     7441 gagaaatacc tgggcctgca aatcgaccct tggaccggca ttgccaggag caacatcagc
     7501 tctaaactcg atagctggct ggagcggatt aatcaggctc cacttaagcc cctccaaaag
     7561 ctggacatcc tgaagactta cacaatccct cggctgacct atatggttga tcacagcgag
     7621 atgaaggctg gggcactcga ggctctggac ttgcagatta gatcagctgt gaaagactgg
     7681 ctccatcttc catcatgtac atgcgatgca atcctgtacg tgagcaccaa agatggcggt
     7741 ctgggagtga ctaaattggc cgggctcatt ccttctatcc aggcacgtag gctgcacagg
     7801 attgctcaga gcccagacga aacgatgaag gcgttcctcg acaaggagca gatggaaaaa
     7861 cagtacgcta aattgtgggt tcaggcaggg gggaaacgtg agaaaatacc ttcaatctgg
     7921 gatgcactgc caacccccgt cttgctgaca acctccgata cactttccga gtgggaagcc
     7981 cctaatccca aaagtaagta tccacgcccg tgcaattggc gccgcaaaga gtttgaaaag
     8041 tggacaaagc tccagtgtca gggtagaggt atccagaatt ttaaagggga cgtcatcagc
     8101 aataattgga ttcagaacta tcgcaggatc ccacatcgca agctcctcac agccgtgcag
     8161 ctccgcgcca acgtctaccc aactagggaa tttttgggcc ggggcagggg tgatgactgt
     8221 gtgaagtttt gccgacactg tgaagtggac ctggagactt gtggccatat catatcatac
     8281 tgtccagtga caaaagaggc ccgaattaag cggcataata gaatttgcga acggctgatt
     8341 gaagaagccg agaaaaagga ctgggtggtc tttaaagaac ctcacattag ggatgcagtc
     8401 aaggaactct ttaagcccga cctgatattt gttaaagagg acagagctct ggtggttgac
     8461 gttaccgtac gtttcgaagc cacaaccact agcctcgaag aagccgcgat cgaaaaggtg
     8521 gataaataca aaaggctgga aacagaagtc cgctccctga caaatgccaa ggacgttctc
     8581 ttcatggggt ttcctcttgg tgcgcggggg aagtggtacc agggtaactt taaactcctc
     8641 gacatgctgg gccttagcga gtctagacaa gtgaccgttg ccaagaccct gagtaccgat
     8701 gccctgattt catccgttga tatcgtacac atgtttgcct caaaggcacg gaagatgaat
     8761 ctcgtaacag tgtctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagcccaag
     8821 aagaagagga aagtcagcgg ggggtctggt ggatctggag gacccgcggc taagagggtg
     8881 aagcttgacg gtggataacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     8941 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     9001 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag aaaattgcat
     9061 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     9121 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     9181 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     9241 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     9301 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     9361 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     9421 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     9481 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     9541 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     9601 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     9661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     9721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     9781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     9841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     9901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     9961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    10021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    10081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    10141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    10201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    10261 ggtctgacac tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    10321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    10381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    10441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    10501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    10561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    10621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    10681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
    10741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    10801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    10861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    10921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    10981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    11041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    11101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    11161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    11221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    11281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    11341 tttagaaaaa taaacaaata ggggttccgc gca
//