LOCUS       sequence-447665-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with MKKCMN 
            amino acids at positions 1135,1136,1218,1219,1335,T1337 with C-term 
            BPNLS-P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Silverstein RA, Kim N, Kroell AS, Walton RT, Delano J, Butcher RM, 
            Pacesa M, Smith BK, Christie KA, Ha LL, Meis RJ, Clark AB, Spinner 
            AD, Lazzarotto CR, Li Y, Matsubara A, Urbina EO, Dahl GA, Correia 
            BE, Marks DS, Tsai SQ, Pinello L, De Ravin SS, Liu Q, Kleinstiver BP
  TITLE     Custom CRISPR-Cas9 PAM variants via scalable engineering and machine
            learning.
  JOURNAL   Nature. 2025 Apr 22. doi: 10.1038/s41586-025-09021-y.
  PUBMED    40262634
REFERENCE   2  (bases 1 to 8451)
  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: "Nature. 
            2025 Apr 22. doi: 10.1038/s41586-025-09021-y."
COMMENT     Sequence Label: pCMV-T7-SpCas9(MKKCMN)-P2A-EGFP (AHK129)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(77..94)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(248..836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(328..347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1007..1798,1799..1867))
                     /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     1630..1649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1868..1972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2051..2070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2242..2621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2622..2825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2776..2796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2867..2886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2867..2885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2893..2902
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2899..7002
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFMKPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKCLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKMYNSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7057..7077
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7090..7155
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7174..7230
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(7231..7233,7234..7236,7237..7950)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7231..7233,7234..7236,7237..7947)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7537..7556)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7884..7905
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7956..7973
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7996..8013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8002..8226
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8297..8313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8297..8313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8310..8332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8321..8337
                     /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(8345..8351,8352..8369,8370..8375))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8390..8411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
       61 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      121 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      181 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      421 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      661 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1081 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1141 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1201 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1261 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1321 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1381 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1441 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1501 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1561 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1621 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1681 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1741 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1801 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1861 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1921 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1981 cccgaaaagt gccacctgac gtcgacggat cgggagatcg atctcccgat cccctagggt
     2041 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     2101 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     2161 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     2221 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     2281 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2341 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2401 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2461 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2521 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2581 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2641 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2701 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     2761 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2821 gagctggttt agtgaaccgt cagatccgct agagatccgc ggccgctaat acgactcact
     2881 atagggagag ccgccaccat ggacaagaag tacagcatcg gcctggacat cggcaccaac
     2941 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     3001 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     3061 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     3121 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     3181 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     3241 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     3301 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     3361 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     3421 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     3481 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     3541 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     3601 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     3661 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     3721 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     3781 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     3841 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     3901 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     3961 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     4021 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     4081 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     4141 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     4201 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     4261 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     4321 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     4381 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     4441 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     4501 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     4561 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     4621 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     4681 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     4741 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     4801 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     4861 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     4921 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     4981 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     5041 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     5101 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     5161 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     5221 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     5281 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     5341 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     5401 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     5461 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     5521 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     5581 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     5641 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     5701 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     5761 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     5821 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     5881 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     5941 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     6001 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     6061 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     6121 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     6181 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     6241 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     6301 atgaagccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     6361 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     6421 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     6481 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     6541 gcctctgcca agtgcctgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     6601 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     6661 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     6721 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     6781 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     6841 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     6901 atgtacaaca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     6961 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acggatccgg cggaggcgga
     7021 agcgggaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtctcg
     7081 agcggaggcg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     7141 gatgacgatg acaagggagg cggtggaagc ggcgcaacaa acttctctct gctgaaacaa
     7201 gccggagatg tcgaagagaa tcctggaccg atggtgagca agggcgagga gctgttcacc
     7261 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     7321 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     7381 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     7441 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     7501 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     7561 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     7621 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     7681 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     7741 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     7801 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     7861 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     7921 actctcggca tggacgagct gtacaagtga ccggtcatca tcaccatcac cattgagttt
     7981 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     8041 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     8101 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     8161 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     8221 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
     8281 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     8341 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
     8401 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac c
//
