LOCUS       sequence-171050-        9291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes gRNA for 3' target of human MIER3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9291)
  AUTHORS   Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts 
            BS, Mackiewicz M, Mendenhall EM, Myers RM
  TITLE     CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins.
  JOURNAL   Genome Res. 2015 Sep 9.
  PUBMED    26355004
REFERENCE   2  (bases 1 to 9291)
  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: "Genome Res.
            2015 Sep 9."
COMMENT     Sequence Label: PX458_MIER3_2
FEATURES             Location/Qualifiers
     source          1..9291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      35..623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     524..543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        685..933
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     685..705
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     856..875
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        955..1030
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1127..1412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1414..1691
                     /label=chicken beta-actin promoter
     intron          1692..1919
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1931..1940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1940..2005
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2012..2032
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2057..6157
                     /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             2057..6157
                     /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             6158..6205
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6221..6274
                     /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             join(6275..6277,6278..6988)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6275..6277,6278..6988)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6578..6597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6925..6946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7016..7033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7022..7229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7238..7378
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7238..7367
                     /label=AAV2 ITR
     rep_origin      7453..7908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7540..7559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7750..7771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7925..7944)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8044..8066
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8104..8122)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8190..8294
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8295..8363,8364..9155)
                     /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(8513..8532)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
       61 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      661 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
      721 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      781 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      841 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      901 gctttatata tcttgtggaa aggacgaaac accgcactct gagtgacctg agtggtttta
      961 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     1021 gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     1081 tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt acataactta
     1141 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
     1201 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     1261 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     1321 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
     1381 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     1441 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     1501 tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg
     1561 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     1621 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     1681 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     1741 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     1801 gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg
     1861 ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg
     1921 ttggaccggt gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat
     1981 tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca
     2041 cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt
     2101 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     2161 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     2221 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     2281 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     2341 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     2401 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     2461 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     2521 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     2581 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     2641 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     2701 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     2761 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     2821 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     2881 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     2941 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     3001 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     3061 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     3121 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     3181 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     3241 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     3301 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     3361 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     3421 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     3481 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     3541 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     3601 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     3661 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     3721 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     3781 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     3841 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     3901 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     3961 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     4021 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     4081 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     4141 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     4201 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     4261 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     4321 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     4381 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     4441 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     4501 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     4561 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     4621 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     4681 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     4741 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     4801 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     4861 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     4921 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     4981 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     5041 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     5101 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     5161 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     5221 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     5281 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     5341 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     5401 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     5461 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     5521 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     5581 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     5641 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     5701 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     5761 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     5821 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     5881 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     5941 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     6001 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     6061 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     6121 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
     6181 ggccggccag gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct
     6241 aacatgcggt gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac
     6301 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6361 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6421 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6481 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     6541 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     6601 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6661 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     6721 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     6781 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     6841 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     6901 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     6961 cactctcggc atggacgagc tgtacaagga attctaacta gagctcgctg atcagcctcg
     7021 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     7081 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     7141 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     7201 tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg
     7261 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     7321 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     7381 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     7441 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     7501 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     7561 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     7621 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     7681 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7741 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     7801 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     7861 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     7921 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     7981 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     8041 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     8101 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     8161 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     8221 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     8281 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     8341 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     8401 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     8461 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     8521 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     8581 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     8641 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     8701 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     8761 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     8821 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     8881 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     8941 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     9001 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     9061 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     9121 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     9181 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     9241 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga g
//
