LOCUS       sequence-393133-        9279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA for TBPL1 KO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9279)
  AUTHORS   Kwan JZJ, Nguyen TF, Uzozie AC, Budzynski MA, Cui J, Lee JMC, Van 
            Petegem F, Lange PF, Teves SS
  TITLE     RNA Polymerase II transcription independent of TBP in murine 
            embryonic stem cells.
  JOURNAL   Elife. 2023 Mar 30;12:e83810. doi: 10.7554/eLife.83810.
  PUBMED    36995326
REFERENCE   2  (bases 1 to 9279)
  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: "2023-03-30"; volume: "12:e83810. doi"; pages: " 
            10.7554/eLife.83810"
COMMENT     Sequence Label: TBPL1 KO 1
FEATURES             Location/Qualifiers
     source          1..9279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2012..2059
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2063..2116
                     /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             2123..2833
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2268..2286)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2858..2875)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2864..3071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3080..3220
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3080..3209
                     /label=AAV2 ITR
     rep_origin      3295..3750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3382..3401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3592..3613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3767..3786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3886..3908
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3946..3964)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4032..4136
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4137..4205,4206..4997)
                     /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(4355..4374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5168..5756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5657..5676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5818..6066
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5818..5838
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5989..6008
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6088..6163
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        6260..6545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        6547..6824
                     /label=chicken beta-actin promoter
     intron          6825..7052
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      7064..7073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7073..7138
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7145..7165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7190..9279,1..2011)
                     /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             join(7190..9279,1..2011)
                     /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"
ORIGIN
        1 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
       61 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
      121 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
      181 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
      241 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
      301 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
      361 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
      421 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
      481 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
      541 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
      601 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
      661 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
      721 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
      781 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
      841 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
      901 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
      961 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     1021 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     1081 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     1141 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     1201 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     1261 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     1321 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     1381 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     1441 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     1501 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     1561 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     1621 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     1681 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     1741 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     1801 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     1861 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     1921 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     1981 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
     2041 ccaggcaaaa aagaaaaagc ttgagggcag aggaagtctg ctaacatgcg gtgacgtgga
     2101 ggagaatccc ggccctgcta gcatggtgag caagggcgag gaggataaca tggccatcat
     2161 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     2221 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2281 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     2341 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     2401 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     2461 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     2521 cttcccctca gacggccccg taatgcagaa gaaaaccatg ggctgggagg cctcctccga
     2581 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     2641 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     2701 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     2761 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     2821 gctgtacaag taagaattcc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     2881 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2941 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3001 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg
     3061 catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3121 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3181 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3241 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3301 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3361 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3421 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3481 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3541 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3601 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3661 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3721 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3781 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3841 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3901 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3961 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4021 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4081 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4141 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4201 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4261 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4321 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4381 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4441 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4501 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4561 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4621 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4681 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4741 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4801 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg
     4861 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4921 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4981 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5041 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5101 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5161 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5221 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5281 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5341 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5401 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5461 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5521 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5581 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5641 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5701 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5761 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag
     5821 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     5881 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     5941 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     6001 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     6061 aacaccgatc actgtctgca tccattggtt ttagagctag aaatagcaag ttaaaataag
     6121 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
     6181 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
     6241 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6301 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
     6361 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6421 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
     6481 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6541 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
     6601 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
     6661 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
     6721 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     6781 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     6841 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     6901 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     6961 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
     7021 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
     7081 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
     7141 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
     7201 cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     7261 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     7321 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     7381 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     7441 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     7501 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     7561 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     7621 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     7681 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     7741 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     7801 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     7861 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     7921 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     7981 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     8041 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     8101 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     8161 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     8221 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     8281 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     8341 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     8401 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     8461 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     8521 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     8581 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     8641 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     8701 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     8761 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     8821 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     8881 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     8941 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     9001 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     9061 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     9121 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     9181 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     9241 cgacggcttc gccaacagaa acttcatgca gctgatcca
//
