LOCUS       Exported               10582 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Plasmid encoding Cas9-53BP1-DN1S fusion with BbsI restriction sites 
            for insertion of guide RNA sequence..
ACCESSION   .
VERSION     .
KEYWORDS    pX458M-53BP1-DN1S
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10582)
  AUTHORS   Jayavaradhan R, Pillis DM, Goodman M, Zhang F, Zhang Y, Andreassen 
            PR, Malik P
  TITLE     CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and 
            inhibits NHEJ specifically at Cas9 target sites.
  JOURNAL   Nat Commun. 2019 Jun 28;10(1):2866. doi: 10.1038/s41467-019-10735-7.
  PUBMED    31253785
REFERENCE   2  (bases 1 to 10582)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-06-28"; volume: "10(1):2866. doi"; pages: " 
            10.1038/s41467-019-10735-7"
FEATURES             Location/Qualifiers
     source          1..10582
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        450..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        737..1014
                     /label=chicken beta-actin promoter
     intron          1015..1242
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1254..1263
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1263..1328
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1335..1355
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1380..5480
                     /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             6765..6812
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6828..6881
                     /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             6882..7595
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             6882..7595
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6924..6945)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7185..7204)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7532..7553
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7623..7640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7629..7836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7845..7985
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7845..7974
                     /label=AAV2 ITR
     rep_origin      8060..8515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8147..8166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8357..8378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8532..8551)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8651..8673
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8711..8729)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8797..8901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8902..9762
                     /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(9120..9139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9933..10521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10422..10441
                     /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 ggtcttcgag aagacctgtt taagagctat gctggaaaca gcatagcaag
      301 tttaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
      361 ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa
      421 ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct
      481 ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      781 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      841 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      901 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      961 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     1021 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     1081 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     1141 tgtaattagc tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg
     1201 tttaattacc tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca
     1261 tggactataa ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg
     1321 acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg
     1381 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
     1441 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     1501 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     1561 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     1621 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1681 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1741 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1801 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     1861 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     1921 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     1981 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     2041 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     2101 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     2161 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     2221 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     2281 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     2341 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     2401 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     2461 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     2521 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     2581 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     2641 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2701 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2761 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     2821 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     2881 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     2941 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     3001 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     3061 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     3121 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     3181 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     3241 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     3301 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     3361 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     3421 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     3481 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     3541 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     3601 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3661 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3721 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3781 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     3841 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
     3901 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     3961 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     4021 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     4081 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     4141 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     4201 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     4261 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     4321 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     4381 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     4441 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     4501 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     4561 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     4621 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4681 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     4741 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4801 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     4861 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     4921 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     4981 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     5041 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     5101 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     5161 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     5221 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     5281 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     5341 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     5401 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     5461 tgtctcagct gggaggcgac acagggtcca caggatccac aggcagcaca gggagcatgg
     5521 gaccacatgg ccatgtctta catcgtcaca tgagaacaat ccgggaagta cgcacacttg
     5581 tcactcgtgt cattacagat gtgtattatg tggatggaac agaagtagaa agaaaagtaa
     5641 ctgaggagac tgaagagcca attgtagagt gtcaggagtg tgaaactgaa gtttcccctt
     5701 cacagactgg gggctcctca ggtgacctgg gggatatcag ctccttctcc tccaaggcat
     5761 ccagcttaca ccgcacatca agtgggacaa gtctctcagc tatgcacagc agtggaagct
     5821 cagggaaagg agccggacca ctcagaggga aaaccagcgg gacagaaccc gcagattttg
     5881 ccttacccag ctcccgagga ggcccaggaa aactgagtcc tagaaaaggg gtcagtcaga
     5941 cagggacgcc agtgtgtgag gaggatggtg atgcaggcct tggcatcaga cagggaggga
     6001 aggctccagt cacgcctcgt gggcgtgggc gaaggggccg cccaccttct cggaccactg
     6061 gaaccagaga aacagctgtg cctggcccct tgggcataga ggacatttca cctaacttgt
     6121 caccagatga taaatccttc agccgtgtcg tgccccgagt gccagactcc accagacgaa
     6181 cagatgtggg tgctggtgct ttgcgtcgta gtgactctcc agaaattcct ttccaggctg
     6241 ctgctggccc ttctgatggc ttagatgcct cctctccagg aaatagcttt gtagggctcc
     6301 gtgttgtagc caagtggtca tccaatggct acttttactc tgggaaaatc acacgagatg
     6361 tcggagctgg gaagtataaa ttgctctttg atgatgggta cgaatgtgat gtgttgggca
     6421 aagacattct gttatgtgac cccatcccgc tggacactga agtgacggcc ctctcggagg
     6481 atgagtattt cagtgcagga gtggtgaaag gacataggaa ggagtctggg gaactgtact
     6541 acagcattga aaaagaaggc caaagaaagt ggtataagcg aatggctgtc atcctgtcct
     6601 tggagcaagg aaacagactg agagagcagt atgggcttgg cccctatgaa gcagtaacac
     6661 ctcttacaaa ggcagcagat atcagcttag acaatttggt ggaagggaag cggaaacgga
     6721 gatctaacgt cagctcccca gccaccccta ctgcctcctc gagcaaaagg ccggcggcca
     6781 cgaaaaaggc cggccaggca aaaaagaaaa aggaattcgg cagtggagag ggcagaggaa
     6841 gtctgctaac atgcggtgac gtcgaggaga atcctggccc agtgagcaag ggcgaggagc
     6901 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     6961 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     7021 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     7081 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     7141 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     7201 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     7261 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     7321 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     7381 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     7441 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     7501 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     7561 ccgggatcac tctcggcatg gacgagctgt acaaggaatt ctaactagag ctcgctgatc
     7621 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     7681 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     7741 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     7801 ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat
     7861 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     7921 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     7981 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     8041 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     8101 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     8161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     8221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     8281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     8401 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     8461 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     8521 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     8581 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8641 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     8701 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     8761 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     8821 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     8881 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     8941 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     9001 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     9061 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     9121 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     9181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     9241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     9301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     9361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     9421 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     9481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     9541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9601 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     9661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     9721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     9781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     9841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     9901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     9961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    10021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    10081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    10141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    10201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    10261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    10321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    10381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    10441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    10501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    10561 tgctggcctt ttgctcacat gt
//
