LOCUS       sequence-177667-        8989 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human codon optimized RecA protein was fused to eSpCas9(1.1) for 
            enhanced genome editing efficiency.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8989)
  AUTHORS   Lin L, Petersen TS, Jensen KT, Bolund L, Kuhn R, Luo Y
  TITLE     Fusion of SpCas9 to E. coli Rec A protein enhances CRISPR-Cas9 
            mediated gene knockout in mammalian cells.
  JOURNAL   J Biotechnol. 2017 Mar 1;247:42-49. doi: 
            10.1016/j.jbiotec.2017.02.024.
  PUBMED    28259533
REFERENCE   2  (bases 1 to 8989)
  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: "J 
            Biotechnol."; date: "2017-03-1"; volume: "247:42-49. doi"; pages: " 
            10.1016/j.jbiotec.2017.02.024"
COMMENT     Sequence Label: P823_eSpCas9(1.1)-RecA
FEATURES             Location/Qualifiers
     source          1..8989
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             5..70
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             77..97
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(122..2659,2660..2662,2663..3124,3125..3127,3128..3295,
                     3296..3298,3299..4222)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4223..4270
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5369..5389
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5414..5563
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(5597..5614)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5603..5810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5819..5959
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5819..5948
                     /label=AAV2 ITR
     rep_origin      6034..6489
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6121..6140)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6331..6352
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6506..6525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6625..6647
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6685..6703)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6771..6875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6876..6944,6945..7736)
                     /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(7094..7113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7907..8495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8396..8415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8557..8805
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8557..8577
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8728..8747
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8824..8899
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
       61 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
      121 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
      181 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
      241 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
      301 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
      361 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
      421 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
      481 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
      541 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
      601 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
      661 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
      721 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
      781 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
      841 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
      901 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
      961 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     1021 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     1081 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     1141 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     1201 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     1261 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     1321 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     1381 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     1441 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     1501 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     1561 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     1621 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     1681 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     1741 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     1801 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     1861 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     1921 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     1981 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     2041 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     2101 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     2161 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     2221 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     2281 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     2341 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     2401 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     2461 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     2521 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     2581 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     2641 cgtgcctcag agctttctgg ccgacgactc catcgacaac aaggtgctga ccagaagcga
     2701 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     2761 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     2821 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     2881 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     2941 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     3001 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     3061 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     3121 ccctgcgctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     3181 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     3241 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaaggcgcc
     3301 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     3361 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     3421 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     3481 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     3541 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     3601 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     3661 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     3721 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     3781 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     3841 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     3901 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     3961 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     4021 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     4081 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     4141 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     4201 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     4261 aaagaaaaag ggcggcggag gatccgaatt cggcggaggc ggcagcatgg ccatcgacga
     4321 gaacaagcag aaggccctgg ccgctgccct gggccagatc gagaagcagt ttggcaaggg
     4381 cagcatcatg cggctgggcg aggacagatc catggacgtg gaaaccatca gcaccggcag
     4441 cctgagcctg gatatcgctc tgggagctgg cggcctgcct atgggcagaa tcgtggaaat
     4501 ctacggcccc gagagcagcg gcaagaccac cctgacactg caagtgatcg ctgccgccca
     4561 gcgggaaggc aagacctgcg cctttatcga tgccgagcac gccctggacc ccatctacgc
     4621 cagaaagctg ggagtggaca tcgacaacct gctgtgcagc cagcccgata caggcgaaca
     4681 ggccctggaa atctgcgacg ccctggctag atctggcgcc gtggatgtga tcgtggtgga
     4741 ttctgtggcc gccctgaccc ccaaggccga gattgagggc gagatcggcg atagccacat
     4801 gggcctggcc gccagaatga tgagccaggc catgaggaag ctggccggca acctgaagca
     4861 gagcaacacc ctgctgatct tcatcaacca gatccgcatg aagatcggcg tgatgttcgg
     4921 caaccccgag acaaccacag gcggcaacgc cctgaagttc tacgcctccg tgcggctgga
     4981 catcagacgg atcggagccg tgaaagaggg ggagaacgtc gtgggcagcg agacaagagt
     5041 gaaggtcgtg aagaacaaga tcgccgctcc cttcaagcag gccgagttcc agatcctgta
     5101 cggcgagggc atcaacttct acggcgagct ggtggacctg ggagtgaaag agaagctgat
     5161 tgagaaggct ggcgcctggt acagctacaa gggcgagaag atcggacagg gcaaggccaa
     5221 tgccaccgcc tggctgaagg acaaccccga aaccgccaaa gaaatcgaga agaaagtgcg
     5281 cgagctgctg ctgagcaacc ccaacagcac ccccgacttc agcgtggacg attctgaggg
     5341 cgtggccgag acaaacgagg acttcccacc caagaaaaag cggaaagtct gagaattctg
     5401 tacaggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct
     5461 cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat
     5521 gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact aagaattcct
     5581 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5641 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5701 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5761 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag
     5821 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     5881 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5941 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6001 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     6061 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     6121 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6181 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6241 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6301 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6361 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     6421 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     6481 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6541 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6601 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6661 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6721 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6841 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6901 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6961 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7021 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7081 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7141 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7201 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7261 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7321 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7381 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7441 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7501 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7561 ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc
     7621 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7681 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7741 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7801 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7861 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7921 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7981 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8041 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8101 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8161 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8221 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8281 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8341 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8401 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8461 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8521 ggttcctggc cttttgctgg ccttttgctc acatgtgagg gcctatttcc catgattcct
     8581 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     8641 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     8701 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     8761 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgggtc ttcgagaaga
     8821 cctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     8881 agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt aaaataaggc
     8941 tagtccgttt ttagcgcgtg cgccaattct gcagacaaat ggctctaga
//
