LOCUS       sequence-260818-        9289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes gRNA for 3' target of human THRA_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9289)
  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 9289)
  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_THRA_iso1_1
FEATURES             Location/Qualifiers
     source          1..9289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..263
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     15..35
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     186..205
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        283..358
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        455..740
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        742..1019
                     /label=chicken beta-actin promoter
     intron          1020..1247
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1259..1268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1268..1333
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1340..1360
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1385..5485
                     /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             1385..5485
                     /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             5486..5533
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5549..5602
                     /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(5603..5605,5606..6316)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5603..5605,5606..6316)
                     /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(5906..5925)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6253..6274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6344..6361)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6350..6557
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6566..6706
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6566..6695
                     /label=AAV2 ITR
     rep_origin      6781..7236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6868..6887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7078..7099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7253..7272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7372..7394
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7432..7450)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7518..7622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7623..7691,7692..8483)
                     /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(7841..7860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8654..9242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9143..9162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca
       61 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
      121 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      181 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      241 tcttgtggaa aggacgaaac accggaagtc taaagcctca gggttttaga gctagaaata
      301 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      361 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
      421 gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc
      481 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
      541 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      601 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      661 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      721 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      781 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      841 tgggggcggg gggggggggg gggsgcgcgc caggcggggc ggggcggggc gaggggcggg
      901 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      961 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     1021 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
     1081 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
     1141 cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat
     1201 taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc
     1261 caccatggac tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga
     1321 cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc
     1381 agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     1441 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     1501 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     1561 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     1621 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     1681 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     1741 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     1801 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     1861 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     1921 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     1981 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     2041 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     2101 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     2161 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     2221 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     2281 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     2341 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     2401 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     2461 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     2521 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     2581 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     2641 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     2701 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     2761 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     2821 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     2881 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     2941 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     3001 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     3061 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     3121 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     3181 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     3241 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     3301 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     3361 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     3421 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     3481 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     3541 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     3601 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     3661 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     3721 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     3781 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     3841 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     3901 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     3961 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     4021 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     4081 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     4141 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     4201 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     4261 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     4321 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     4381 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     4441 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     4501 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     4561 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     4621 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     4681 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     4741 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     4801 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     4861 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     4921 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     4981 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     5041 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     5101 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     5161 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     5221 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     5281 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     5341 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     5401 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     5461 cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc
     5521 aaaaaagaaa aaggaattcg gcagtggaga gggcagagga agtctgctaa catgcggtga
     5581 cgtcgaggag aatcctggcc cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     5641 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     5701 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     5761 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     5821 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     5881 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     5941 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     6001 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     6061 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     6121 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     6181 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     6241 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     6301 ggacgagctg tacaaggaat tctaactaga gctcgctgat cagcctcgac tgtgccttct
     6361 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     6421 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     6481 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     6541 agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     6601 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     6661 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     6721 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     6781 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6841 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6901 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6961 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     7021 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7081 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     7141 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     7201 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     7261 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     7321 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     7381 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     7441 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     7501 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7561 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7621 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7681 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7741 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7801 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7861 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7921 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7981 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     8041 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     8101 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     8161 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     8221 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     8281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     8341 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     8401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     8461 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     8521 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8581 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8641 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8701 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8761 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8821 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8881 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8941 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     9001 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     9061 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     9121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     9181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     9241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcc
//
