LOCUS       sequence-233476-        8519 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 8519)
  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 8519)
  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..8519
                     /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"
     promoter        4555..4832
                     /label=chicken beta-actin promoter
     intron          4833..5061
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5073..5082
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5082..5147
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5154..5174
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5199..7736,7737..7739,7740..8201,8202..8204,8205..8372,
                     8373..8375,8376..8519,1..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
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     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 gaccaatctg ggagcccctg ccgccttcaa gtactttgac
      661 accaccatcg 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 gccagcgccc 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 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     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 gtccttctag 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 caccggtcag gattctcggt
     4081 aagaagttta 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 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
     4741 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     4801 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc
     4861 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     4921 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
     4981 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
     5041 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg
     5101 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
     5161 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
     5221 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     5281 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     5341 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     5401 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     5461 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     5521 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     5581 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     5641 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     5701 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     5761 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     5821 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     5881 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     5941 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     6001 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     6061 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     6121 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     6181 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     6241 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     6301 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     6361 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     6421 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     6481 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     6541 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     6601 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     6661 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     6721 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     6781 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     6841 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     6901 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     6961 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     7021 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     7081 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     7141 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     7201 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     7261 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     7321 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     7381 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     7441 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     7501 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     7561 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     7621 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     7681 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctggccg
     7741 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     7801 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     7861 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     7921 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     7981 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     8041 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     8101 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     8161 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc tgcgctggaa agcgagttcg
     8221 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     8281 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     8341 agattaccct ggccaacggc gagatccgga aggcgcctct gatcgagaca aacggcgaaa
     8401 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     8461 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagag
//
