LOCUS       sequence-238800-        8518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA targeting mouse Gsk3b and eSpCas9 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8518)
  AUTHORS   Ochiai H, Hayashi T, Umeda M, Yoshimura M, Harada A, Shimizu Y, 
            Nakano K, Saitoh N, Liu Z, Yamamoto T, Okamura T, Ohkawa Y, Kimura 
            H, Nikaido I
  TITLE     Genome-wide kinetic properties of transcriptional bursting in mouse 
            embryonic stem cells.
  JOURNAL   Sci Adv. 2020 Jun 17;6(25):eaaz6699. doi: 10.1126/sciadv.aaz6699. 
            eCollection 2020 Jun.
  PUBMED    32596448
REFERENCE   2  (bases 1 to 8518)
  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: "Sci Adv."; 
            date: "2020-06-17"; volume: "6(25):eaaz6699. doi"; pages: " 
            10.1126/sciadv.aaz6699. eCollection 2020 Jun"
COMMENT     Sequence Label: eSpCas9-EF-Gsk3b(new)
FEATURES             Location/Qualifiers
     source          1..8518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             781..828
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(856..873)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    862..1069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1078..1218
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1078..1207
                     /label=AAV2 ITR
     rep_origin      1293..1748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1380..1399)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1590..1611
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1765..1784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1884..1906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1944..1962)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2030..2134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2135..2203,2204..2995)
                     /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(2353..2372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3166..3754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3655..3674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3816..4064
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3816..3836
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3987..4006
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        4268..4553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          4833..5060
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5072..5081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5081..5146
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5153..5173
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5198..7735,7736..7738,7739..8200,8201..8203,8204..8371,
                     8372..8374,8375..8518,1..780)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDDTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLINLGAPAAFKYFDTIIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
       61 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
      121 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
      181 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
      241 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
      301 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
      361 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
      421 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
      481 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
      541 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
      601 atcatccacc tgtttaccct gatcaatctg ggagcccctg ccgccttcaa gtactttgac
      661 accatcatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
      721 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
      781 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagta agaattccta
      841 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
      901 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
      961 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1021 aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag cggccgcagg
     1081 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     1141 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     1201 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     1261 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     1321 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     1381 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1441 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1501 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1561 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1621 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     1681 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     1741 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     1801 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1861 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1921 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1981 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2041 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2101 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2161 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2221 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2281 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2341 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2401 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2461 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2521 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2581 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2641 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2701 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2761 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2821 tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc
     2881 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2941 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3001 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3061 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3301 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3781 gttcctggcc ttttgctggc cttttgctca catgtgaggg cctatttccc atgattcctt
     3841 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
     3901 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     3961 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     4021 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgactcc acctgaacag
     4081 cccccgttta agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta
     4141 tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag
     4201 ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag
     4261 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4321 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4381 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4441 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
     4501 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     4561 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     4621 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
     4681 cgccrggsgg gggggggggg gggggggggg gggggggggg gggggggggg ggggcggcgg
     4741 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     4801 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     4861 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     4921 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
     4981 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
     5041 ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg
     5101 agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa
     5161 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
     5221 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     5281 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     5341 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     5401 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     5461 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     5521 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     5581 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     5641 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     5701 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     5761 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     5821 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     5881 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     5941 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     6001 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     6061 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     6121 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     6181 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     6241 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     6301 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacgaca ccgaggaact
     6361 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     6421 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     6481 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     6541 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     6601 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     6661 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     6721 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     6781 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     6841 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     6901 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     6961 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     7021 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     7081 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     7141 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     7201 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     7261 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     7321 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     7381 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     7441 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     7501 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     7561 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     7621 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     7681 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctggccga
     7741 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     7801 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     7861 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     7921 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     7981 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     8041 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     8101 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     8161 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct gcgctggaaa gcgagttcgt
     8221 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     8281 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     8341 gattaccctg gccaacggcg agatccggaa ggcgcctctg atcgagacaa acggcgaaac
     8401 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     8461 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagag
//
