LOCUS       sequence-171034-        9290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes gRNA for 3' target of human MBD1_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9290)
  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 9290)
  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_MBD1_iso1_2
FEATURES             Location/Qualifiers
     source          1..9290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      65..653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     554..573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        715..963
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     715..735
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     886..905
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        984..1059
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1156..1441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1443..1720
                     /label=chicken beta-actin promoter
     intron          1721..1948
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1960..1969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1969..2034
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2041..2061
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2086..6186
                     /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             2086..6186
                     /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             6187..6234
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6250..6303
                     /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(6304..6306,6307..7017)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6304..6306,6307..7017)
                     /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(6607..6626)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6954..6975
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7045..7062)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7051..7258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7267..7407
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7267..7396
                     /label=AAV2 ITR
     rep_origin      7482..7937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7569..7588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7779..7800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7954..7973)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8073..8095
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8133..8151)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8219..8323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8324..8392,8393..9184)
                     /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(8542..8561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
       61 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      181 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtgagggc
      721 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
      781 ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
      841 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
      901 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
      961 accgctagaa agcagtagac tgggttttag agctagaaat agcaagttaa aataaggcta
     1021 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa
     1081 atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat
     1141 ggctctagag gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1201 cgacccccgc ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg
     1261 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1321 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     1501 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     1561 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
     1621 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     1681 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     1741 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     1801 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag
     1861 caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga
     1921 gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac
     1981 cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc
     2041 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc
     2101 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     2161 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     2221 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     2281 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     2341 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     2401 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     2461 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     2521 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     2581 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     2641 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     2701 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     2761 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     2821 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     2881 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     2941 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     3001 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     3061 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     3121 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     3181 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     3241 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     3301 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     3361 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     3421 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     3481 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     3541 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     3601 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     3661 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     3721 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     3781 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     3841 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     3901 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     3961 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     4021 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     4081 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     4141 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     4201 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     4261 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     4321 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     4381 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     4441 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     4501 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     4561 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     4621 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     4681 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     4741 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     4801 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     4861 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     4921 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     4981 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     5041 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     5101 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     5161 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     5221 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     5281 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     5341 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     5401 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     5461 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     5521 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     5581 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     5641 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     5701 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     5761 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     5821 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     5881 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     5941 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     6001 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     6061 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     6121 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     6181 ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaaggaattc
     6241 ggcagtggag agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc
     6301 ccagtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     6361 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     6421 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     6481 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6541 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     6601 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     6661 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     6721 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     6781 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     6841 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     6901 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     6961 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggaa
     7021 ttctaactag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     7081 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     7141 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     7201 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
     7261 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     7321 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     7381 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     7441 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     7501 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     7561 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     7621 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     7681 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7741 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7801 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     7861 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     7921 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7981 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     8041 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     8101 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     8161 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     8221 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     8281 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     8341 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     8401 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     8461 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     8521 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     8581 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     8641 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     8701 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8761 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8821 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8881 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8941 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     9001 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     9061 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     9121 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     9181 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     9241 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
//
