LOCUS       sequence-393128-        9720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA for HSF2 KO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9720)
  AUTHORS   Price RM, Budzynski MA, Shen J, Mitchell JE, Kwan JZJ, Teves SS
  TITLE     Heat shock transcription factors demonstrate a distinct mode of 
            interaction with mitotic chromosomes.
  JOURNAL   Nucleic Acids Res. 2023 Apr 28:gkad304. doi: 10.1093/nar/gkad304.
  PUBMED    37114996
REFERENCE   2  (bases 1 to 9720)
  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: "Nucleic 
            Acids Res. 2023 Apr 28:gkad304. doi: 10.1093/nar/gkad304."
COMMENT     Sequence Label: HSF2 KO
FEATURES             Location/Qualifiers
     source          1..9720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      60..648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     549..568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        710..958
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        980..1055
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1151..1399
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        1421..1496
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1593..1878
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1880..2157
                     /label=chicken beta-actin promoter
     intron          2158..2385
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2397..2406
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2406..2471
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2478..2498
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2523..6623
                     /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             2523..6623
                     /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             6624..6671
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6675..6728
                     /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             6735..7445
                     /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(6880..6898)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(7470..7487)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7476..7683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7692..7832
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7692..7821
                     /label=AAV2 ITR
     rep_origin      7907..8362
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7994..8013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8204..8225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8379..8398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8498..8520
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8558..8576)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8644..8748
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8749..8817,8818..9609)
                     /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(8967..8986)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
       61 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      121 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      181 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
      241 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      301 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      361 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      421 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      481 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      541 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      601 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      661 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
      721 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      781 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      841 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      901 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccga
      961 attctactac cgaacgcggg ttttagagct agaaatagca agttaaaata aggctagtcc
     1021 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg
     1081 ttgtacaaaa aagcaggctt taaaggaacc aattcagtcg actggatccg gtaccaaggt
     1141 cgggcaggaa gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     1201 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     1261 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     1321 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     1381 gtggaaagga cgaaacaccg tccaagccag gtaggaatgg gttttagagc tagaaatagc
     1441 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     1501 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     1561 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
     1621 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     1681 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1741 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1801 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1861 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     1921 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     1981 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
     2041 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     2101 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     2161 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     2221 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     2281 ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta
     2341 atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca
     2401 ccatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
     2461 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
     2521 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     2581 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     2641 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     2701 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     2761 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     2821 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2881 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2941 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     3001 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     3061 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     3121 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     3181 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     3241 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     3301 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     3361 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     3421 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     3481 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     3541 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     3601 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     3661 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3721 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     3781 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3841 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3901 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3961 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     4021 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     4081 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     4141 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     4201 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     4261 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     4321 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     4381 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     4441 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     4501 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     4561 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     4621 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     4681 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     4741 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     4801 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     4861 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     4921 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     4981 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     5041 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     5101 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     5161 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     5221 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     5281 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     5341 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     5401 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     5461 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     5521 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     5581 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     5641 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     5701 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     5761 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     5821 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     5881 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     5941 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     6001 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     6061 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     6121 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     6181 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     6241 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     6301 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     6361 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     6421 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     6481 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     6541 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     6601 acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa
     6661 aaaagaaaaa gcttgagggc agaggaagtc tgctaacatg cggtgacgtg gaggagaatc
     6721 ccggccctgc tagcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     6781 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6841 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     6901 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     6961 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     7021 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     7081 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     7141 cagacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt
     7201 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7261 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     7321 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     7381 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     7441 agtaagaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     7501 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     7561 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     7621 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     7681 ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     7741 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     7801 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     7861 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     7921 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     7981 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     8041 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8101 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     8161 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     8221 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     8281 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     8341 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     8401 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     8461 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     8521 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     8581 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     8641 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     8701 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     8761 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     8821 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     8881 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     8941 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     9001 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     9061 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     9121 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     9181 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     9241 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     9301 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     9361 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     9421 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     9481 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     9541 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     9601 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     9661 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
//
