LOCUS       sequence-196972-        8992 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA/SpCas9 expression plasmid with Cas9 VRER mutations (NGCG PAM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8992)
  AUTHORS   Lambrus BG, Moyer TC, Holland AJ
  TITLE     Applying the auxin-inducible degradation system for rapid protein 
            depletion in mammalian cells.
  JOURNAL   Methods Cell Biol. 2018;144:107-135. doi: 
            10.1016/bs.mcb.2018.03.004. Epub 2018 Apr 5.
  PUBMED    29804665
REFERENCE   2  (bases 1 to 8992)
  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: "Methods 
            Cell Biol."; date: "2018"; volume: "144:107-135. doi"; pages: " 
            10.1016/bs.mcb.2018.03.004. Epub 2018 Apr 5"
COMMENT     Sequence Label: px459 VRER
FEATURES             Location/Qualifiers
     source          1..8992
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      232..241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             241..306
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             313..333
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(358..3756,3757..3759,3760..4005,4006..4008,4009..4356,
                     4357..4359,4360..4362,4363..4365,4366..4458)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/G1218R/R1335E/T1337R mutations"
                     /label=Cas9 VRER
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4459..4506
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4522..4575
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4576..5175
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4576..4595)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5072..5092
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5203..5220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5209..5416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5425..5565
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5425..5554
                     /label=AAV2 ITR
     rep_origin      5640..6095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5727..5746)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5937..5958
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6112..6131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6231..6253
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6291..6309)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6377..6481
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6482..6550,6551..7342)
                     /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     complement(6700..6719)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7513..8101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8002..8021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8163..8411
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8163..8183
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8334..8353
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8430..8505
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8602..8887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
       61 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      121 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
      181 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
      241 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
      301 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
      361 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
      421 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      481 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      541 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      601 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      661 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      721 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
      781 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
      841 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
      901 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
      961 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     1021 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     1081 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     1141 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     1201 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1261 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     1321 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1381 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1441 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1501 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1561 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     1621 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1681 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1741 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     1801 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     1861 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     1921 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     1981 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     2041 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     2101 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     2161 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat agtgttgact
     2221 cttaccctct ttgaagatcg ggaaatgatt gaggaaagac taaaaacata cgctcacctg
     2281 ttcgacgata aggttatgaa acagttaaag aggcgtcgct atacgggctg gggacgattg
     2341 tcgcggaaac ttatcaacgg gataagagac aagcaaagtg gtaaaactat tctcgatttt
     2401 ctaaagagcg acggcttcgc caataggaac tttatgcagc tgatccatga tgactcttta
     2461 accttcaaag aggatataca aaaggcacag gtttccggac aaggggactc attgcacgaa
     2521 catattgcga atcttgctgg ttcgccagcc atcaaaaagg gcatactcca gacagtcaaa
     2581 gtagtggatg agctagttaa ggtcatggga cgtcacaaac cggaaaacat tgtaatcgag
     2641 atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa acagtcgaga gcggatgaag
     2701 agaatagaag agggtattaa agaactgggc agccagatct taaaggagca tcctgtggaa
     2761 aatacccaat tgcagaacga gaaactttac ctctattacc tacaaaatgg aagggacatg
     2821 tatgttgatc aggaactgga cataaaccgt ttatctgatt acgacgtcga tcacattgta
     2881 ccccaatcct ttttgaagga cgattcaatc gacaataaag tgcttacacg ctcggataag
     2941 aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg taaagaaaat gaagaactat
     3001 tggcggcagc tcctaaatgc gaaactgata acgcaaagaa agttcgataa cttaactaaa
     3061 gctgagaggg gtggcttgtc tgaacttgac aaggccggat ttattaaacg tcagctcgtg
     3121 gaaacccgcc aaatcacaaa gcatgttgca cagatactag attcccgaat gaatacgaaa
     3181 tacgacgaga acgataagct gattcgggaa gtcaaagtaa tcactttaaa gtcaaaattg
     3241 gtgtcggact tcagaaagga ttttcaattc tataaagtta gggagataaa taactaccac
     3301 catgcgcacg acgcttatct taatgccgtc gtagggaccg cactcattaa gaaatacccg
     3361 aagctagaaa gtgagtttgt gtatggtgat tacaaagttt atgacgtccg taagatgatc
     3421 gcgaaaagcg aacaggagat aggcaaggct acagccaaat acttctttta ttctaacatt
     3481 atgaatttct ttaagacgga aatcactctg gcaaacggag agatacgcaa acgaccttta
     3541 attgaaacca atggggagac aggtgaaatc gtatgggata agggccggga cttcgcgacg
     3601 gtgagaaaag ttttgtccat gccccaagtc aacatagtaa agaaaactga ggtgcagacc
     3661 ggagggtttt caaaggaatc gattcttcca aaaaggaata gtgataagct catcgctcgt
     3721 aaaaaggact gggacccgaa aaagtacggt ggcttcgtga gccctacagt tgcctattct
     3781 gtcctagtag tggcaaaagt tgagaaggga aaatccaaga aactgaagtc agtcaaagaa
     3841 ttattgggga taacgattat ggagcgctcg tcttttgaaa agaaccccat cgacttcctt
     3901 gaggcgaaag gttacaagga agtaaaaaag gatctcataa ttaaactacc aaagtatagt
     3961 ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta gcgccagaga gcttcaaaag
     4021 gggaacgaac tcgcactacc gtctaaatac gtgaatttcc tgtatttagc gtcccattac
     4081 gagaagttga aaggttcacc tgaagataac gaacagaagc aactttttgt tgagcagcac
     4141 aaacattatc tcgacgaaat catagagcaa atttcggaat tcagtaagag agtcatccta
     4201 gctgatgcca atctggacaa agtattaagc gcatacaaca agcacaggga taaacccata
     4261 cgtgagcagg cggaaaatat tatccatttg tttactctta ccaacctcgg cgctccagcc
     4321 gcattcaagt attttgacac aacgatagat cgcaaagagt acagatctac caaggaggtg
     4381 ctagacgcga cactgattca ccaatccatc acgggattat atgaaactcg gatagatttg
     4441 tcacagcttg ggggtgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     4501 aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag
     4561 gagaatcctg gcccaatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4621 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4681 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4741 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4801 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4861 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4921 caccggccca aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag
     4981 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5041 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5101 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5161 aagcccggtg cctgagaatt ctaactagag ctcgctgatc agcctcgact gtgccttcta
     5221 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5281 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5341 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata
     5401 gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     5461 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     5521 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     5581 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     5641 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5701 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5761 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5821 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     5881 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5941 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     6001 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6061 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     6121 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     6181 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6241 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     6301 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6361 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6421 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6481 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6541 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6601 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6661 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6721 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6781 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6841 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6901 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6961 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7021 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7081 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7141 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     7201 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7261 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7321 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7381 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7441 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7501 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7561 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7621 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7681 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7741 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7801 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7861 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7921 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7981 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8041 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8101 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8161 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     8221 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     8281 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     8341 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     8401 gacgaaacac cgggtcttcg agaagacctg ttttagagct agaaatagca agttaaaata
     8461 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
     8521 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
     8581 acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc
     8641 gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg
     8701 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     8761 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca
     8821 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     8881 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     8941 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gg
//
