LOCUS       sequence-383855-        8395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 nickase expression vector bearing a sgRNA targeting Exon 2 of 
            human SOX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  AUTHORS   Landshammer A, Bolondi A, Kretzmer H, Much C, Buschow R, Rose A, Wu 
            HJ, Mackowiak SD, Braendl B, Giesselmann P, Tornisiello R, Parsi KM,
            Huey J, Mielke T, Meierhofer D, Maehr R, Hnisz D, Michor F, Rinn JL,
            Meissner A
  TITLE     T-REX17 is a transiently expressed non-coding RNA essential for 
            human endoderm formation.
  JOURNAL   Elife. 2023 Jan 31;12:e83077. doi: 10.7554/eLife.83077.
  PUBMED    36719724
REFERENCE   2  (bases 1 to 8395)
  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: "Elife."; 
            date: "2023-01-31"; volume: "12:e83077. doi"; pages: " 
            10.7554/eLife.83077"
COMMENT     Sequence Label: pX335_U6-Chimeric_BB-CBh-hSpCas9n(D10A)_SOX17_...
FEATURES             Location/Qualifiers
     source          1..8395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          122..349
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             370..396
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             400..420
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(430..453,454..456,457..4530)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             430..4530
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4534..4554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4591..4608)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4597..4804
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4813..4953
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4813..4942
                     /label=AAV2 ITR
     rep_origin      5028..5483
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5115..5134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5325..5346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5500..5519)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5619..5641
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5679..5697)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5765..5869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5870..5938,5939..6730)
                     /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(6088..6107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6901..7489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7390..7409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7551..7799
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7551..7571
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7722..7741
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7820..7895
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7992..8277
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
       61 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      121 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      181 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      241 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
      301 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
      361 gccaccatgt acccatacga tgttccagat tacgcttcgc cgaagaaaaa gcgcaaggtc
      421 gaagcgtccg acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg
      481 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
      541 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
      601 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
      661 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
      721 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
      781 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
      841 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
      901 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
      961 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     1021 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     1081 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     1141 ggcaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     1201 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     1261 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     1321 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     1381 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     1441 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     1501 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     1561 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     1621 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     1681 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     1741 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     1801 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     1861 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     1921 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     1981 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     2041 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     2101 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     2161 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     2221 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     2281 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     2341 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     2401 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     2461 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     2521 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     2581 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     2641 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     2701 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     2761 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     2821 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     2881 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     2941 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     3001 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     3061 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     3121 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     3181 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     3241 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     3301 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     3361 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     3421 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     3481 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     3541 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     3601 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     3661 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     3721 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     3781 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     3841 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     3901 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     3961 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     4021 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     4081 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     4141 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     4201 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     4261 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     4321 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     4381 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     4441 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     4501 cggatcgacc tgtctcagct gggaggcgac agccccaaga agaagagaaa ggtggaggcc
     4561 agctaagaat tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     4621 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4681 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4741 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     4801 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4861 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4921 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4981 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     5041 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5101 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5161 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5221 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     5281 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5341 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     5401 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5461 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5521 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5581 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5641 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5701 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5761 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5821 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5881 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5941 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6001 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6061 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6121 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6181 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6241 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6301 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6361 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6421 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6481 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6541 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     6601 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6661 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6721 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6841 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6901 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6961 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7021 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7081 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7141 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7201 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7261 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7321 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7381 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7441 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7501 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
     7561 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     7621 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     7681 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     7741 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     7801 cagtaatata ccgcggagtg ttttagagct agaaatagca agttaaaata aggctagtcc
     7861 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag
     7921 caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct
     7981 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     8041 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
     8101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     8161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
     8221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     8281 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     8341 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggg
//
