LOCUS       sequence-447628-        8611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCBh Human expression plasmid for SpCas9 enzyme with MRRWMR amino 
            acids at positions 1135,1136,1218,1219,1335,T1337 with C-term 
            BPNLS-P2A-mTagBFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  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 8611)
  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: pCBh-T7-SpCas9(MRRWMR)-P2A-mTagBFP2(RAS3579)
FEATURES             Location/Qualifiers
     source          1..8611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(155..946,947..1015))
                     /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     778..797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1016..1120)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1199..1218)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1404..1689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1691..1967
                     /label=chicken beta-actin promoter
     intron          1968..2195
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2207..2216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2213..6316
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFMRPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARWLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKMYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6371..6391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6404..6469
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6488..6544
                     /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             6545..7255
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             7264..7281
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7304..7321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7310..7534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7605..7621)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7605..7621)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7618..7640)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7629..7645
                     /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(7653..7659,7660..7677,7678..7683))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7698..7719
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7836..7853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8007..8595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8087..8106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
       61 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      121 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      181 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      241 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      301 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      361 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      421 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      481 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      541 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      601 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      661 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
      721 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
      781 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
      841 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
      901 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
      961 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1141 cacctgacgt cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta
     1201 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     1261 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     1321 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     1381 atacgcgtct agatcagggt acccgttaca taacttacgg taaatggccc gcctggctga
     1441 ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac tttccattga
     1501 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     1561 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc
     1621 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     1681 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     1741 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     1801 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
     1861 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1921 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1981 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     2041 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     2101 agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt
     2161 acctggagca cctgcctgaa atcacttttt ttcaggttgg accggcgcca ccatggacaa
     2221 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
     2281 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     2341 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     2401 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     2461 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     2521 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     2581 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     2641 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     2701 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     2761 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     2821 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     2881 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     2941 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     3001 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     3061 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     3121 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     3181 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     3241 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     3301 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     3361 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     3421 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     3481 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     3541 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     3601 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     3661 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     3721 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     3781 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     3841 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     3901 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     3961 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     4021 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     4081 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     4141 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     4201 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     4261 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     4321 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     4381 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     4441 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     4501 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     4561 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     4621 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     4681 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     4741 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     4801 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     4861 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     4921 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     4981 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     5041 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     5101 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     5161 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     5221 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     5281 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     5341 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     5401 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     5461 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     5521 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     5581 gaaggactgg gaccctaaga agtacggcgg cttcatgaga cccaccgtgg cctattctgt
     5641 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     5701 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     5761 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     5821 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccagatggc tgcagaaggg
     5881 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     5941 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     6001 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     6061 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     6121 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     6181 cttcaagtac tttgacacca ccatcgaccg gaagatgtac agaagcacca aagaggtgct
     6241 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     6301 tcagctggga ggcgacggat ccggcggagg cggaagcggg aaaagaaccg ccgacggcag
     6361 cgaattcgag cccaagaaga agaggaaagt ctcgagcgga ggcgactaca aagaccatga
     6421 cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg gaggcggtgg
     6481 aagcggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
     6541 accgatggtg tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat
     6601 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta
     6661 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
     6721 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg
     6781 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     6841 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct
     6901 catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa
     6961 gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga
     7021 aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa
     7081 gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt
     7141 ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga
     7201 ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaaac ttaattgacc
     7261 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     7321 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     7381 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     7441 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     7501 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     7561 ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7621 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7681 aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7741 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7801 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7861 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7921 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7981 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8041 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8101 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8161 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8221 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8281 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8341 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8401 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8461 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8521 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8581 aaaaaaggat ctcaagaaga tcctttgatc t
//
