LOCUS       sequence-177023-        8505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses high specificity SpCas9. Px330-like plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8505)
  AUTHORS   Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F
  TITLE     Rationally engineered Cas9 nucleases with improved specificity.
  JOURNAL   Science. 2015 Dec 1. pii: aad5227.
  PUBMED    26628643
REFERENCE   2  (bases 1 to 8505)
  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: "Science. 
            2015 Dec 1. pii: aad5227."
COMMENT     Sequence Label: eSpCas9(1.1)
FEATURES             Location/Qualifiers
     source          1..8505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          172..399
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      411..420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             420..485
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             492..512
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(537..3074,3075..3077,3078..3539,3540..3542,3543..3710,
                     3711..3713,3714..4637)
                     /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             4638..4685
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4713..4730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4719..4926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4935..5075
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4935..5064
                     /label=AAV2 ITR
     rep_origin      5150..5605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5237..5256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5447..5468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5622..5641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5741..5763
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5801..5819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5887..5991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5992..6060,6061..6852)
                     /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(6210..6229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7023..7611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7512..7531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7673..7921
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7673..7693
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7844..7863
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7940..8015
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8112..8397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8399..8505,1..171)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
       61 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
      121 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      181 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      241 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      301 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
      361 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
      421 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
      481 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
      541 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
      601 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
      661 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
      721 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
      781 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
      841 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
      901 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
      961 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     1021 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     1081 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     1141 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     1201 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     1261 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     1321 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     1381 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     1441 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     1501 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     1561 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     1621 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     1681 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     1741 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     1801 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     1861 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     1921 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     1981 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     2041 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     2101 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     2161 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     2221 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     2281 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     2341 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     2401 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     2461 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     2521 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     2581 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     2641 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     2701 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     2761 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     2821 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     2881 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     2941 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     3001 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     3061 ctcagagctt tctggccgac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     3121 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     3181 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     3241 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     3301 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     3361 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     3421 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     3481 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtaccctg
     3541 cgctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     3601 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     3661 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag gcgcctctga
     3721 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     3781 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     3841 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     3901 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     3961 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     4021 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     4081 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     4141 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     4201 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     4261 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     4321 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     4381 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     4441 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     4501 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     4561 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     4621 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     4681 aaaagtaaga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4741 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4801 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4861 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     4921 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4981 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     5041 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     5101 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5221 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5341 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5461 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     5521 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5581 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5641 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5701 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5761 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
     6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtgagggcct
     7681 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
     7741 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     7801 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     7861 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     7921 cgggtcttcg agaagacctg ttttagagct agaaatagca agttaaaata aggctagtcc
     7981 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag
     8041 caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct
     8101 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     8161 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
     8221 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     8281 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
     8341 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     8401 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     8461 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcg
//
