LOCUS       sequence-388911-        9720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding for 2 gRNAs targeting the human TERT gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9720)
  AUTHORS   Laprade H, Querido E, Smith MJ, Guerit D, Crimmins H, Conomos D, 
            Pourret E, Chartrand P, Sfeir A
  TITLE     Single-Molecule Imaging of Telomerase RNA Reveals a 
            Recruitment-Retention Model for Telomere Elongation.
  JOURNAL   Mol Cell. 2020 Jul 2;79(1):115-126.e6. doi: 
            10.1016/j.molcel.2020.05.005. Epub 2020 Jun 3.
  PUBMED    32497497
REFERENCE   2  (bases 1 to 9720)
  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"; doi: 
            "10.1016/j.molcel.2020.05.005"; journalName: "Mol Cell"; date: 
            "2020-07-2- 2"; volume: "79"; issue: "1"; pages: "115-126.e6"
COMMENT     Sequence Label: pSpCas9_gRNA1TERTKO_BsagRNA5tertko_2A-GFP
FEATURES             Location/Qualifiers
     source          1..9720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        437..685
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        706..781
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        872..1157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1437..1664
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1676..1685
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1685..1750
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1757..1777
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1802..5902
                     /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             1802..5902
                     /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             5903..5950
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5966..6019
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6020..6022,6023..6733)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6020..6022,6023..6733)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6323..6342)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6670..6691
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6761..6778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6767..6974
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6983..7123
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6983..7112
                     /label=AAV2 ITR
     rep_origin      7198..7653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7285..7304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7495..7516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7670..7689)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7789..7811
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7849..7867)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7935..8039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8040..8108,8109..8900)
                     /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(8258..8277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9071..9659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9560..9579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg cttcggggtc cactagcgtg gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagagagg gcctatttcc catgattcct tcatatttgc atatacgata
      481 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
      541 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      601 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      661 tatcttgtgg aaaggacgaa acaccgcgca gctacctgcc caacagtttt agagctagaa
      721 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      781 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      841 tgcgccaatt ctgcagacaa atggcggtac ccgttacata acttacggta aatggcccgc
      901 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt
      961 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1021 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     1081 attgtgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     1141 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     1201 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     1261 gggcgggggg gggggggggg cgcgcgccgg gggggggggg gggggggggg ggggggggsg
     1321 gggsgrggsg grgaggkgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
     1381 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
     1441 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
     1501 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     1561 gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa
     1621 tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac
     1681 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
     1741 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
     1801 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
     1861 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     1921 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     1981 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     2041 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     2101 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     2161 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     2221 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     2281 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     2341 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     2401 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     2461 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     2521 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     2581 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     2641 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     2701 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     2761 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     2821 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     2881 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     2941 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     3001 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     3061 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     3121 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     3181 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     3241 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     3301 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     3361 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     3421 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     3481 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     3541 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     3601 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     3661 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     3721 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     3781 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     3841 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     3901 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     3961 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     4021 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     4081 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     4141 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     4201 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     4261 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     4321 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     4381 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     4441 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     4501 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     4561 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     4621 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     4681 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     4741 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     4801 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     4861 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     4921 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     4981 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     5041 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     5101 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     5161 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     5221 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     5281 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     5341 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     5401 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     5461 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     5521 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     5581 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     5641 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     5701 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     5761 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     5821 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     5881 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     5941 aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
     6001 cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     6061 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     6121 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     6181 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     6241 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     6301 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     6361 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     6421 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     6481 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     6541 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     6601 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     6661 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     6721 cgagctgtac aaggaattct aactagagct cgctgatcag cctcgactgt gccttctagt
     6781 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6841 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6901 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc
     6961 aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     7021 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     7081 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     7141 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     7201 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     7261 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     7321 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     7381 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     7441 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     7501 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     7561 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     7621 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     7681 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     7741 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     7801 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     7861 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     7921 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     7981 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     8041 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     8101 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     8161 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     8221 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     8281 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     8341 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     8401 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     8461 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     8521 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     8581 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     8641 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     8701 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc
     8761 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     8821 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     8881 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     8941 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     9001 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     9061 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     9121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     9181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     9241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     9301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     9361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     9421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     9481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     9541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     9601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     9661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
