LOCUS       sequence-202454-        8637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses R1335K mutant SpCas9 fused to FKBP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8637)
  AUTHORS   Scot Wolfe
  TITLE     Inducible Cas9-DNA-binding-domain fusions for precise genome editing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8637)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCSD_SpCas9MT3-FKBP_SL
FEATURES             Location/Qualifiers
     source          1..8637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..55
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     37..54
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    110..134
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      138..147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             174..4274
                     /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
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKKYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(4323..4325,4326..4646)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    4688..4822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4712..4731)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4766..4785
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4935..4955)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4935..4953)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4974..4990)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4974..4990)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4987..5009)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4998..5014
                     /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(5022..5028,5029..5046,5047..5052))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5067..5088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5205..5222)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5376..5964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5456..5475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6135..6926,6927..6995))
                     /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     6758..6777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6996..7100)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7126..7581)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7263..7284)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7475..7494
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(7655..8637,1..2)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca
       61 agctacttgt tctttttgca ggatcccatc gattcgaatt caaggatcaa caagtttgta
      121 caaaaaagca ggctggcgcc accatggccg ggactagtgg gcgtcccggg tccgacaaga
      181 agtacagcat cggcctggat atcggtacca acagcgtcgg ctgggccgtc attacggacg
      241 agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa
      301 agaagaacct cattggagcc ctcctgttcg actccgggga gacggccgaa gccacgcggc
      361 tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg
      421 agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt
      481 cctttttggt ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg
      541 acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagctggtag
      601 acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcac atgatcaaat
      661 ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac
      721 tctttatcca actggttcag acttacaatc agcttttcga ggagaacccg atcaacgcat
      781 ccggcgttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa
      841 acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc
      901 tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc
      961 tgcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg
     1021 accagtacgc agaccttttt ttggcggcaa agaacctgtc agacgccatt ctgctgagtg
     1081 atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc
     1141 gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc
     1201 ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg
     1261 acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg
     1321 acggcaccga ggagctgctg gtaaagctga acagagaaga tctgttgcgc aaacagcgca
     1381 ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca
     1441 ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc
     1501 tcacatttcg gataccctac tatgtaggcc ccctcgctcg gggaaattcc agattcgcgt
     1561 ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata
     1621 agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta
     1681 acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc
     1741 tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc
     1801 agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc
     1861 tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg
     1921 aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca
     1981 aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta
     2041 cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg
     2101 acgacaaagt catgaaacag ctcaagagac gccgatatac aggatggggg cggctgtcaa
     2161 gaaaactgat caatggcatc cgagacaagc agagtggaaa gacaatcctg gattttctta
     2221 agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct
     2281 ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca
     2341 tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg
     2401 tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg
     2461 cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga
     2521 ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca
     2581 cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg
     2641 tggatcagga actggacatc aaccggttgt ccgactacga cgtggatcat atcgtgcccc
     2701 aaagctttct caaagatgat tctattgata ataaagtgtt gacaagatcc gataaaaata
     2761 gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc
     2821 ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg
     2881 aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga
     2941 cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg
     3001 atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct
     3061 cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg
     3121 cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc
     3181 tggaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa
     3241 agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga
     3301 attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg
     3361 aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc
     3421 gcaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag
     3481 gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa
     3541 aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac
     3601 tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc
     3661 tgggcatcac aatcatggag cgatccagct tcgagaaaaa ccccatcgac tttctcgaag
     3721 cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gctgcccaag tactctctct
     3781 ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta
     3841 acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa
     3901 agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac
     3961 actaccttga tgagatcatc gagcaaataa gcgagttctc caaaagagtg atcctcgccg
     4021 acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg
     4081 agcaggcaga aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct
     4141 tcaagtactt cgacaccacc atagacagaa agaagtacac ctctacaaag gaggtcctgg
     4201 acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc
     4261 agctcggtgg agacggcacc ggcgggccta gggcgggctc cagggggagt gggagctccg
     4321 gaatgggagt gcaggtggaa accatctccc caggagacgg gcgcaccttc cccaagcgcg
     4381 gccagacctg cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa tttgattcct
     4441 ctcgggacag aaacaagccc tttaagttta tgctaggcaa gcaggaggtg atccgaggct
     4501 gggaagaagg ggttgcccag atgagtgtgg gtcagagagc caaactgact atatctccag
     4561 attatgccta tggtgccact gggcacccag gcatcatccc accacatgcc actctcgtct
     4621 tcgatgtgga gcttctaaaa ctggaactcg agtaataatc tagaactata gtgagtcgta
     4681 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4741 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4801 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4861 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     4921 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4981 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5041 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5101 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5161 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5221 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5281 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5341 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5401 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5461 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5521 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5581 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5641 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5701 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5761 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5821 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5881 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5941 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6001 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6061 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6121 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6181 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6241 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6301 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6361 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6421 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6481 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6541 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6601 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6661 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6721 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6781 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6841 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6901 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6961 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7021 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7081 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     7141 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     7201 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     7261 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     7321 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     7381 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     7441 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     7501 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     7561 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     7621 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca atatggcgta
     7681 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
     7741 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
     7801 cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg ggaggggtct
     7861 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
     7921 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
     7981 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
     8041 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
     8101 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
     8161 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
     8221 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
     8281 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
     8341 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
     8401 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag gacgccaggg
     8461 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
     8521 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
     8581 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttaggga
//
