LOCUS       sequence-328916-       12774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of drug-inducible dSpyCas9-mCherry-APEX2 for 
            biotin-labeling genomic element-associated protein factors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12774)
  AUTHORS   Gao XD, Tu LC, Mir A, Rodriguez T, Ding Y, Leszyk J, Dekker J, 
            Shaffer SA, Zhu LJ, Wolfe SA, Sontheimer EJ
  TITLE     C-BERST: defining subnuclear proteomic landscapes at genomic 
            elements with dCas9-APEX2.
  JOURNAL   Nat Methods. 2018 Jun;15(6):433-436. doi: 10.1038/s41592-018-0006-2.
            Epub 2018 May 7.
  PUBMED    29735996
REFERENCE   2  (bases 1 to 12774)
  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"; doi: 
            "10.1038/s41592-018-0006-2"; journalName: "Nat Methods"; date: 
            "2018-06"; volume: "15"; issue: "6"; pages: "433-436"
COMMENT     Sequence Label: pEJS578_DD-dSpyCas9-mCherry-APEX2
FEATURES             Location/Qualifiers
     source          1..12774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2965..2974
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2977..2997
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3004..3051
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3064..3384
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KPE"
     CDS             join(3409..3432,3433..3435,3436..5919,5920..5925,5926..5991,
                     5992..5994,5995..7509)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(3409..3432,3433..3435,3436..5922,5923..5925,
                     5926..7509)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3409..7509
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7528..7548
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7561..8268
                     /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(7706..7724)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7974..7993
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8296..8319
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8320..9066
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             9073..9093
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9100..9147
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    9164..9752
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9217..9237)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9635..9646)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9827..10060
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10179..10197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10265..10369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10370..10438,10439..11230)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10588..10607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11401..11989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11890..11909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12143..12160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12400..12421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12436..12441,12442..12459,12460..12466)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12474..12490
                     /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)."
     primer_bind     12479..12501
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12498..12514
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12498..12514
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12564..12583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggtt aaggtgaacg cgtggccacc atggccccta aaaagaaacg caaggtcgaa
     3001 gataaacgcc cagcagctac aaagaaggca ggtcaagcca agaaaaagaa ggaagatgca
     3061 tgcggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc
     3121 cagacctgtg tggtgcacta caccgggatg cttgaagatg gaaagaaagt cgattcctcc
     3181 cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg
     3241 gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat
     3301 tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc
     3361 gatgtggagc ttctaaaacc ggaaggcagt gaattcggaa gcggaagcga caagaagtac
     3421 tccattgggc tcgctatcgg taccaacagc gtcggctggg ccgtcattac ggacgagtac
     3481 aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag
     3541 aacctcattg gagccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa
     3601 agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc
     3661 tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt
     3721 ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag
     3781 gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct ggtagacagt
     3841 actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcacatgat caaatttcgg
     3901 ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt
     3961 atccaactgg ttcagactta caatcagctt ttcgaggaga acccgatcaa cgcatccggc
     4021 gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc
     4081 atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca
     4141 ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagctgcaa
     4201 ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag
     4261 tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt
     4321 ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat
     4381 gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag
     4441 aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc
     4501 ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc
     4561 accgaggagc tgctggtaaa gctgaacaga gaagatctgt tgcgcaaaca gcgcactttc
     4621 gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg
     4681 caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca
     4741 tttcggatac cctactatgt aggccccctc gctcggggaa attccagatt cgcgtggatg
     4801 actcgcaaat cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg
     4861 gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa
     4921 aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc
     4981 aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag
     5041 aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa
     5101 gaagactatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat
     5161 cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac
     5221 ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg
     5281 tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac
     5341 aaagtcatga aacagctcaa gagacgccga tatacaggat gggggcggct gtcaagaaaa
     5401 ctgatcaatg gcatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc
     5461 gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag
     5521 gaggacatcc agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct
     5581 aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat
     5641 gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga
     5701 gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa
     5761 gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag
     5821 cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat
     5881 caggaactgg acatcaaccg gttgtccgac tacgacgtgg atgctatcgt gccccaaagc
     5941 tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg
     6001 aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag
     6061 ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga
     6121 ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc
     6181 cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa
     6241 aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat
     6301 ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat
     6361 gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagctggaa
     6421 tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct
     6481 gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt
     6541 ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca
     6601 aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccgcaag
     6661 gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc
     6721 tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat
     6781 tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt
     6841 gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc
     6901 atcacaatca tggagcgatc cagcttcgag aaaaacccca tcgactttct cgaagcgaaa
     6961 ggatataaag aggtcaaaaa agacctcatc attaagctgc ccaagtactc tctctttgag
     7021 cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag
     7081 ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc
     7141 aaagggtctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac
     7201 cttgatgaga tcatcgagca aataagcgag ttctccaaaa gagtgatcct cgccgacgct
     7261 aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag
     7321 gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag
     7381 tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc
     7441 acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc
     7501 ggtggagacg gctctactag tggctctccc aagaagaaga ggaaggtagg atccggaagt
     7561 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     7621 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     7681 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     7741 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     7801 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     7861 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc actgcaggac
     7921 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     7981 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     8041 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     8101 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     8161 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     8221 cgcgctgagg gccgccactc caccggcggc atggacgagc tgtacaagtc tggcggccgc
     8281 ggcggtggag gtagcgacta caaggatgac gacgataagg gaaagtctta cccaactgtg
     8341 agtgctgatt accaggacgc cgttgagaag gcgaagaaga agctcagagg cttcatcgct
     8401 gagaagagat gcgctcctct aatgctccgt ttggcattcc actctgctgg aacctttgac
     8461 aagggcacga agaccggtgg acccttcgga accatcaagc accctgccga actggctcac
     8521 agcgctaaca acggtcttga catcgctgtt aggcttttgg agccactcaa ggcggagttc
     8581 cctattttga gctacgccga tttctaccag ttggctggcg ttgttgccgt tgaggtcacg
     8641 ggtggaccta aggttccatt ccaccctgga agagaggaca agcctgagcc accaccagag
     8701 ggtcgcttgc ccgatcccac taagggttct gaccatttga gagatgtgtt tggcaaagct
     8761 atggggctta ctgaccaaga tatcgttgct ctatctgggg gtcacactat tggagctgca
     8821 cacaaggagc gttctggatt tgagggtccc tggacctcta atcctcttat tttcgacaac
     8881 tcatacttca cggagttgtt gagtggtgag aaggaaggtc tccttcagct accttctgac
     8941 aaggctcttt tgtctgaccc tgtattccgc cctctcgttg acaaatatgc agcggacgaa
     9001 gatgccttct ttgctgatta cgctgaggct caccaaaagc tttccgagct tgggtttgct
     9061 gatgccctcg agcctaaaaa gaaacgcaag gtcgaagata aacgcccagc agctacaaag
     9121 aaggcaggtc aagccaagaa aaagaagggc tcttaatcta gataatcaac ctctggatta
     9181 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     9241 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     9301 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     9361 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     9421 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     9481 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     9541 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     9601 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     9661 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     9721 gagtcggatc tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac
     9781 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     9841 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     9901 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     9961 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
    10021 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
    10081 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta
    10141 attaaggaaa gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt
    10201 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
    10261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
    10321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
    10381 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
    10441 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
    10501 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
    10561 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
    10621 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
    10681 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
    10741 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
    10801 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
    10861 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
    10921 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
    10981 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
    11041 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
    11101 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
    11161 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
    11221 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
    11281 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
    11341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
    11401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
    11461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
    11521 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
    11581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
    11641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
    11701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
    11761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
    11821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
    11881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
    11941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
    12001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
    12061 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
    12121 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
    12181 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga
    12241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
    12301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag
    12361 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
    12421 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
    12481 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc
    12541 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
    12601 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
    12661 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
    12721 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagt
//
