LOCUS       sequence-419661-       11858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HDR Universal Surrogate Reporter with a sgRNA and consistently 
            expressed SpCas9. The PuroR reporter gene was designed to be 
            repaired as a selecting marker gene only by the sgRNA/Cas9-triggered
            HDR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11858)
  AUTHORS   Yan N, Sun Y, Fang Y, Deng J, Mu L, Xu K, Mymryk JS, Zhang Z
  TITLE     A Universal Surrogate Reporter for Efficient Enrichment of 
            CRISPR/Cas9-Mediated Homology-Directed Repair in Mammalian Cells.
  JOURNAL   Mol Ther Nucleic Acids. 2020 Mar 6;19:775-789. doi: 
            10.1016/j.omtn.2019.12.021. Epub 2019 Dec 24.
  PUBMED    31955009
REFERENCE   2  (bases 1 to 11858)
  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: "Mol Ther 
            Nucleic Acids."; date: "2020-03-6"; volume: "19:775-789. doi"; 
            pages: " 10.1016/j.omtn.2019.12.021. Epub 2019 Dec 24"
COMMENT     Sequence Label: pHDR-USR-SpCas9
FEATURES             Location/Qualifiers
     source          1..11858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     188..207
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        286..361
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        458..743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1023..1250
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1262..1271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1271..1336
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1343..1363
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1388..5488
                     /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             1388..5488
                     /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             5489..5536
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    5570..5777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        5812..6191
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6192..6395
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6346..6366
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        6440..6458
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    7032..7256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        7294..7312
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7327..7920
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     repeat_region   8201..8235
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8726..8742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8726..8742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8739..8761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8750..8766
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8774..8780,8781..8798,8799..8804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8819..8840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8957..8974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9128..9716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9208..9227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9887..10678,10679..10747))
                     /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     10510..10529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10748..10852)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10878..11333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11015..11036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11227..11246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11460..11482
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11474..11491
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11475..11491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11621..11683
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 cgaattcctg cagcccgggg gatcctttcc catgattcct tcatatttgc atatacgata
       61 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
      121 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      181 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      241 tatcttgtgg aaaggacgaa acaccgttct ccgaacgtgt cacgtgtttt agagctagaa
      301 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      361 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      421 tgcgccaatt ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg
      481 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
      541 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      601 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      661 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      721 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      781 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      841 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsggggggs
      901 ggggsggggs grggsggrgr ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      961 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     1021 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     1081 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     1141 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
     1201 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
     1261 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
     1321 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
     1381 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     1441 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     1501 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     1561 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     1621 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     1681 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     1741 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     1801 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     1861 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     1921 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1981 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     2041 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     2101 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     2161 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     2221 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     2281 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     2341 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     2401 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     2461 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     2521 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     2581 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     2641 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     2701 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     2761 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     2821 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     2881 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     2941 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     3001 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     3061 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     3121 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     3181 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     3241 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     3301 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     3361 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     3421 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     3481 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     3541 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     3601 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     3661 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     3721 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     3781 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     3841 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     3901 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     3961 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     4021 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     4081 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     4141 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     4201 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     4261 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     4321 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     4381 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     4441 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     4501 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     4561 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     4621 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     4681 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     4741 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     4801 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     4861 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     4921 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4981 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     5041 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     5101 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     5161 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     5221 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     5281 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     5341 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     5401 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     5461 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     5521 ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat cagcctcgac tgtgccttct
     5581 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5641 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5701 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     5761 agcaggcatg ctggggagcg gccgccgatg tacgggccag atatacgcgt tgacattgat
     5821 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5881 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5941 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6001 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6061 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6121 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6181 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6241 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6301 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6361 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     6421 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcatg
     6481 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     6541 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     6601 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     6661 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggagttc tccgaacgtg
     6721 tcacgtggga ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     6781 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     6841 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     6901 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6961 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gggcccgttt aaacccgctg
     7021 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     7081 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     7141 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     7201 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     7261 tgaggcggaa agaaccagct ggggctcaga tcttaatacg actcactata gggcgaattg
     7321 ggtaccaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     7381 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7441 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7501 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     7561 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     7621 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     7681 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     7741 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     7801 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     7861 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     7921 aagcttgata tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat
     7981 gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat
     8041 gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac
     8101 aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga
     8161 ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag
     8221 actatctttc tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg
     8281 ctcagtcatc gcccaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt
     8341 tcccgcgagg ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac
     8401 gttgagcgaa aacgcacgtt taccatgatg attcgggaag gtgtggccat gcacgccttt
     8461 aacggtgaac tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca
     8521 cagttccgga tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg
     8581 aactgccgtg ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc
     8641 gaggacgggt atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc
     8701 ggcgggcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     8761 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     8821 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     8881 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     8941 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     9001 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     9061 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     9121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     9181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     9241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     9301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     9361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     9421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     9481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     9541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     9601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     9661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     9721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     9781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     9841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     9901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     9961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    10021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    10081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    10141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    10201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    10261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    10321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    10381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    10441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    10501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    10561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    10621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    10681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    10741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    10801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    10861 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
    10921 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
    10981 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
    11041 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
    11101 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
    11161 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
    11221 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
    11281 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
    11341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
    11401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
    11461 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcc tcgttcattc acgtttttga
    11521 acccgtggag gacgggcaga ctcgcggtgc aaatgtgttt tacagcgtga tggagcagat
    11581 gaagatgctc gacacgctgc agaacacgca gctagattaa ccctagaaag ataatcatat
    11641 tgtgacgtac gttaaagata atcatgtgta aaattgacgc atgtgtttta tcggtctgta
    11701 tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt acacttacat
    11761 actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaacaaa
    11821 aactcaaaat ttcttctata aagtaacaaa acttttat
//
