LOCUS       sequence-447688-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with SKNCKS 
            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(SKNCKS)-P2A-EGFP (RAS2951)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2037..2056)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2228..2607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2608..2811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2762..2782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2853..2872
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2853..2871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2879..2888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2885..6988
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFSKPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASANCLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKKYSSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7043..7063
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7076..7141
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7160..7216
                     /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(7217..7219,7220..7222,7223..7936)
                     /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(7217..7219,7220..7222,7223..7933)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7523..7542)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7870..7891
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7942..7959
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7982..7999)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7988..8212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8283..8299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8283..8299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8296..8318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8307..8323
                     /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(8331..8337,8338..8355,8356..8361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8376..8397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac tctcagtaca
     2041 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     2101 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     2161 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     2221 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     2281 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2341 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2401 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2461 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2521 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2581 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2641 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2701 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2761 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     2821 aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag ggagagccgc
     2881 caccatggac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     2941 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     3001 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     3061 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     3121 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     3181 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     3241 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     3301 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     3361 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     3421 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     3481 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     3541 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     3601 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     3661 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     3721 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     3781 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     3841 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     3901 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     3961 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     4021 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     4081 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     4141 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     4201 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     4261 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     4321 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     4381 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     4441 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     4501 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     4561 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     4621 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     4681 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     4741 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     4801 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     4861 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     4921 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     4981 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     5041 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     5101 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     5161 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     5221 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     5281 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     5341 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     5401 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     5461 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     5521 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     5581 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     5641 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     5701 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     5761 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     5821 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     5881 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     5941 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     6001 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     6061 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     6121 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     6181 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     6241 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcagca agcccaccgt
     6301 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     6361 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     6421 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     6481 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccaactg
     6541 cctgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     6601 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     6661 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     6721 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     6781 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     6841 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaagt acagcagcac
     6901 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     6961 gatcgacctg tctcagctgg gaggcgacgg atccggcgga ggcggaagcg ggaaaagaac
     7021 cgccgacggc agcgaattcg agcccaagaa gaagaggaaa gtctcgagcg gaggcgacta
     7081 caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa
     7141 gggaggcggt ggaagcggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga
     7201 agagaatcct ggaccgatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     7261 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     7321 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     7381 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     7441 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     7501 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     7561 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     7621 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     7681 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     7741 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     7801 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     7861 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     7921 cgagctgtac aagtgaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     7981 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     8041 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     8101 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     8161 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     8221 gcggaaagaa ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca
     8281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     8341 gccggaagca taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt
     8401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct g
//
