LOCUS       sequence-447691-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with VKMAKG 
            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(VKMAKG)-P2A-EGFP (RAS3015)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(67..84)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(238..826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(318..337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(997..1788,1789..1857))
                     /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     1620..1639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1858..1962)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2041..2060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2232..2611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2612..2815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2766..2786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2857..2876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2857..2875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2883..2892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2889..6992
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFVKPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAMALQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKKYGSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7047..7067
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7080..7145
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7164..7220
                     /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(7221..7223,7224..7226,7227..7940)
                     /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(7221..7223,7224..7226,7227..7937)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7527..7546)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7874..7895
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7946..7963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7986..8003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7992..8216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8287..8303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8287..8303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8300..8322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8311..8327
                     /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(8335..8341,8342..8359,8360..8365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8380..8401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
       61 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1141 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1801 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1861 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1921 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     1981 gccacctgac gtcgacggat cgggagatcg atctcccgat cccctagggt cgactctcag
     2041 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2101 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2161 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2221 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2281 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2341 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2401 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2461 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2521 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2581 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2641 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2701 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2761 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     2821 agtgaaccgt cagatccgct agagatccgc ggccgctaat acgactcact atagggagag
     2881 ccgccaccat ggacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     2941 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     3001 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     3061 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     3121 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     3181 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     3241 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     3301 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     3361 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     3421 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     3481 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     3541 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     3601 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     3661 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     3721 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     3781 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     3841 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     3901 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     3961 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     4021 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     4081 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     4141 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     4201 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     4261 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     4321 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     4381 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     4441 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     4501 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     4561 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     4621 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     4681 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     4741 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     4801 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     4861 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     4921 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     4981 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     5041 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     5101 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     5161 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     5221 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     5281 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     5341 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     5401 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     5461 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     5521 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     5581 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     5641 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     5701 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     5761 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     5821 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     5881 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     5941 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     6001 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     6061 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     6121 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     6181 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     6241 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gtgaagccca
     6301 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     6361 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     6421 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     6481 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgcca
     6541 tggccctgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     6601 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     6661 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     6721 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     6781 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     6841 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aagtacggca
     6901 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     6961 cacggatcga cctgtctcag ctgggaggcg acggatccgg cggaggcgga agcgggaaaa
     7021 gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtctcg agcggaggcg
     7081 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg
     7141 acaagggagg cggtggaagc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     7201 tcgaagagaa tcctggaccg atggtgagca agggcgagga gctgttcacc ggggtggtgc
     7261 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     7321 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     7381 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     7441 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     7501 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     7561 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     7621 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     7681 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     7741 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     7801 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     7861 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     7921 tggacgagct gtacaagtga ccggtcatca tcaccatcac cattgagttt aaacccgctg
     7981 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     8041 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     8101 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     8161 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     8221 tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga gcttggcgta
     8281 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     8341 acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct aactcacatt
     8401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc a
//
