LOCUS       sequence-255011-        8463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA for targeting mouse Utf1 locus using CRISPR-cas technique.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8463)
  AUTHORS   Benchetrit H, Jaber M, Zayat V, Sebban S, Pushett A, Makedonski K, 
            Zakheim Z, Radwan A, Maoz N, Lasry R, Renous N, Inbar M, Ram O, 
            Kaplan T, Buganim Y
  TITLE     Direct Induction of the Three Pre-implantation Blastocyst Cell Types
            from Fibroblasts.
  JOURNAL   Cell Stem Cell. 2019 Jun 6;24(6):983-994.e7. doi: 
            10.1016/j.stem.2019.03.018. Epub 2019 Apr 25.
  PUBMED    31031139
REFERENCE   2  (bases 1 to 8463)
  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"; doi: 
            "10.1016/j.stem.2019.03.018"; journalName: "Cell Stem Cell"; date: 
            "2019-06-6- 6"; volume: "24"; issue: "6"; pages: "983-994.e7"
COMMENT     Sequence Label: pX330-Utf1 gRNA
FEATURES             Location/Qualifiers
     source          1..8463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..257
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9..29
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     180..199
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        278..353
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        450..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          971..1198
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1210..1219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1219..1284
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1291..1311
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1336..5436
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1336..5436
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5437..5484
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5512..5529)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5518..5725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5734..5874
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5734..5863
                     /label=AAV2 ITR
     rep_origin      5949..6404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6036..6055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6246..6267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6421..6440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6540..6562
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6600..6618)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6686..6790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6791..6859,6860..7651)
                     /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(7009..7028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7822..8410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8311..8330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacatgtga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
       61 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
      121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
      181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
      241 ggaaaggacg aaacaccgcc atccccatct caaacctgtt ttagagctag aaatagcaag
      301 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
      361 ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa
      421 ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct
      481 ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      781 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      841 gcgggggggg gggggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
      901 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      961 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     1021 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     1081 cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt
     1141 tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt
     1201 tggaccggtg ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt
     1261 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
     1321 ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     1381 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     1441 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     1501 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     1561 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     1621 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     1681 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     1741 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     1801 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     1861 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     1921 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     1981 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     2041 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     2101 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     2161 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     2221 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     2281 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     2341 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     2401 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     2461 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     2521 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     2581 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     2641 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     2701 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     2761 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     2821 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     2881 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     2941 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     3001 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     3061 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     3121 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     3181 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     3241 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     3301 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     3361 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     3421 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     3481 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     3541 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     3601 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     3661 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     3721 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     3781 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     3841 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     3901 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     3961 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     4021 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     4081 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     4141 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     4201 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     4261 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     4321 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     4381 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     4441 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     4501 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     4561 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     4621 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     4681 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     4741 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     4801 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     4861 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     4921 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     4981 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     5041 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     5101 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     5161 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     5221 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     5281 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     5341 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     5401 gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag
     5461 gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca gcctcgactg
     5521 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     5581 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     5641 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     5701 aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca
     5761 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     5821 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     5881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     5941 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6001 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     6061 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6121 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     6181 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6241 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     6301 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     6361 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     6421 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     6481 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6541 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     6601 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6661 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6721 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6781 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6841 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6901 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6961 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7021 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7081 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7141 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7201 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7261 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7321 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7381 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7441 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7501 gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7561 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7621 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7681 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7741 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7801 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7861 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7921 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7981 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8041 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8101 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8161 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8221 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8281 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8341 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8401 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8461 tgc
//
