LOCUS       sequence-447660-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with LWQYQH 
            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(LWQYQH)-P2A-EGFP (RAS2885)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(73..90)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(244..832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(324..343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1003..1794,1795..1863))
                     /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     1626..1645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1864..1968)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2047..2066)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2238..2617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2618..2821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2772..2792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2863..2882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2863..2881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2889..2898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2895..6998
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAQYLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYHSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7053..7073
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7086..7151
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7170..7226
                     /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(7227..7229,7230..7232,7233..7946)
                     /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(7227..7229,7230..7232,7233..7943)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7533..7552)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7880..7901
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7952..7969
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7992..8009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7998..8222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8293..8309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8293..8309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8306..8328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8317..8333
                     /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(8341..8347,8348..8365,8366..8371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8386..8407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
       61 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1981 aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac
     2041 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     2101 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     2161 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     2221 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     2281 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2341 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2401 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2461 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2521 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2581 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2641 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2701 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2761 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2821 tggtttagtg aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag
     2881 ggagagccgc caccatggac aagaagtaca gcatcggcct ggacatcggc accaactctg
     2941 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     3001 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     3061 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     3121 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     3181 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     3241 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     3301 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     3361 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     3421 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     3481 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     3541 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     3601 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     3661 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     3721 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     3781 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     3841 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     3901 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     3961 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     4021 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     4081 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     4141 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     4201 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     4261 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     4321 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     4381 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     4441 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     4501 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     4561 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     4621 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     4681 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     4741 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     4801 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     4861 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     4921 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     4981 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     5041 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     5101 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     5161 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     5221 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     5281 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     5341 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     5401 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     5461 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     5521 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     5581 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     5641 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     5701 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     5761 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     5821 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     5881 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     5941 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     6001 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     6061 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     6121 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     6181 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     6241 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcctgt
     6301 ggcccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     6361 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     6421 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     6481 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     6541 ctgcccagta cctgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     6601 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     6661 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     6721 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     6781 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     6841 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagcagt
     6901 accacagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     6961 acgagacacg gatcgacctg tctcagctgg gaggcgacgg atccggcgga ggcggaagcg
     7021 ggaaaagaac cgccgacggc agcgaattcg agcccaagaa gaagaggaaa gtctcgagcg
     7081 gaggcgacta caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg
     7141 acgatgacaa gggaggcggt ggaagcggcg caacaaactt ctctctgctg aaacaagccg
     7201 gagatgtcga agagaatcct ggaccgatgg tgagcaaggg cgaggagctg ttcaccgggg
     7261 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     7321 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     7381 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     7441 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     7501 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     7561 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     7621 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     7681 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     7741 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     7801 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     7861 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     7921 tcggcatgga cgagctgtac aagtgaccgg tcatcatcac catcaccatt gagtttaaac
     7981 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     8041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     8101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     8161 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     8221 ggcttctgag gcggaaagaa ccagctgggg ctcgataccg tcgacctcta gctagagctt
     8281 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8341 caacatacga gccggaagca taaagtgtaa agcctagggt gcctaatgag tgagctaact
     8401 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt c
//
