LOCUS       sequence-334829-       14660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of dCas9-EGFP-HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14660)
  AUTHORS   Wang H, Xu X, Nguyen CM, Liu Y, Gao Y, Lin X, Daley T, Kipniss NH, 
            La Russa M, Qi LS
  TITLE     CRISPR-Mediated Programmable 3D Genome Positioning and Nuclear 
            Organization.
  JOURNAL   Cell. 2018 Nov 15;175(5):1405-1417.e14. doi: 
            10.1016/j.cell.2018.09.013. Epub 2018 Oct 11.
  PUBMED    30318144
REFERENCE   2  (bases 1 to 14660)
  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: 
            "10.1016/j.cell.2018.09.013"; journalName: "Cell"; date: 
            "2018-11-15- 15"; volume: "175"; issue: "5"; pages: "1405-1417.e14"
COMMENT     Sequence Label: pHR-TRE3G-dCas9-EGFP-HaloTag
FEATURES             Location/Qualifiers
     source          1..14660
                     /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    2050..2167
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2229..2578
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2237..2255
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2273..2291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2309..2327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2345..2363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2381..2399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2417..2435
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2453..2471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2517..2541
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2622..2648,2649..2651,2652..5138,5139..5141,
                     5142..6725)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2622..6725
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6729..6755
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6735..6761
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             6774..6794
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6801..6821
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6861..6863,6864..6866,6867..7577)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6861..6863,6864..6866,6867..7577)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7167..7186)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7514..7535
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7596..8483
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     misc_feature    8524..9112
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8577..8597)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8995..9006)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9115..9131)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9116..9132)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9222..9455
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(9480..9498)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(9481..9498)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(9788..9807)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(10021..10040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10140..10162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10200..10218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10286..10390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10391..10459,10460..11251)
                     /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(10609..10628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11422..12010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11911..11930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12164..12181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(12251..12271)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12256..12585
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12436..12571
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          13719..13784
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             13914..13934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    14359..14493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14396..14415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14450..14469
                     /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 tatcgccaaa tggcagtatt
     2041 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2161 aaattttcgg gtttattaca gggacagcag agatccagtt tatcgatgag gccctttcgt
     2221 cttcactcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     2281 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     2341 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     2401 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     2461 agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct
     2521 cgtttagtga accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttac
     2581 gcgtctgcag gatatcaagc ttgcggtacc gcgggcccgg gatggacaag aagtacagca
     2641 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     2701 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     2761 tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg ctgaagagaa
     2821 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     2881 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     2941 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     3001 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     3061 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     3121 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     3181 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     3241 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     3301 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg
     3361 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     3421 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     3481 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     3541 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     3601 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     3661 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc gatggcggag
     3721 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     3781 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     3841 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     3901 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     3961 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     4021 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgcca
     4081 gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     4141 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag
     4201 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaaag
     4261 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     4321 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     4381 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     4441 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     4501 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     4561 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     4621 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     4681 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     4741 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     4801 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     4861 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     4921 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     4981 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     5041 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     5101 aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc
     5161 tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac cggggcaaga
     5221 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cgccagctgc
     5281 tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc gagagaggcg
     5341 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     5401 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaacg
     5461 acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     5521 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     5581 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     5641 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     5701 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     5761 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     5821 gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg cggaaagtgc
     5881 tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     5941 aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag aaggactggg
     6001 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     6061 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     6121 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     6181 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     6241 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     6301 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     6361 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa cactacctgg
     6421 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     6481 tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga gagcaggccg
     6541 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     6601 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     6661 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     6721 gcgacgccta tccctatgac gtgcccgatt atgccagcct gggcagcggc tcccccaaga
     6781 aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacggcatt ggtagtggga
     6841 gcaacggcag cagcggatcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     6901 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     6961 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     7021 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     7081 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     7141 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     7201 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     7261 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     7321 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     7381 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     7441 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     7501 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     7561 tggacgagct gtacaagggt ggtgaaacgc gtggtgcaga aatcggtact ggctttccat
     7621 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
     7681 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
     7741 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
     7801 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
     7861 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
     7921 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
     7981 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
     8041 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
     8101 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
     8161 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
     8221 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
     8281 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
     8341 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     8401 gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc
     8461 tgtcgacgct cgagatttcc ggctagcctg caggcatgca agcttgatat caagcttatc
     8521 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     8581 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     8641 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     8701 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     8761 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     8821 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     8881 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     8941 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     9001 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     9061 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     9121 cgtcgacctc gagggaatta attcgagctc ggtaccttta agaccaatga cttacaaggc
     9181 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     9241 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     9301 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     9361 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     9421 cagacccttt tagtcagtgt ggaaaatctc tagcagcatc tagaattaat tccgtgtatt
     9481 ctatagtgtc acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg
     9541 cgttctaacg acaatatgta caagcctaat tgtgtagcat ctggcttact gaagcagacc
     9601 ctatcatctc tctcgtaaac tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt
     9661 tctggtaacg ccgtcccgca cccggaaatg gtcagcgaac caatcagcag ggtcatcgct
     9721 agccagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     9781 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     9841 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     9901 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     9961 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaatggt gcactctcag
    10021 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
    10081 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
    10141 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
    10201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
    10261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
    10321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    10381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
    10441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
    10501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
    10561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
    10621 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
    10681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
    10741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
    10801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
    10861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
    10921 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
    10981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    11041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    11101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    11161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    11221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    11281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    11341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    11401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    11461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    11521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
    11581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    11641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    11701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    11761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
    11821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
    11881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
    11941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
    12001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
    12061 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
    12121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
    12181 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
    12241 atgcagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
    12301 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
    12361 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
    12421 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
    12481 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
    12541 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tggacacaag
    12601 acaggcttgc gagatatgtt tgagaatacc actttatccc gcgtcaggga gaggcagtgc
    12661 gtaaaaagac gcggactcat gtgaaatact ggtttttagt gcgccagatc tctataatct
    12721 cgcgcaacct attttcccct cgaacacttt ttaagccgta gataaacagg ctgggacact
    12781 tcacatgagc gaaaaataca tcgtcacctg ggacatgttg cagatccatg cacgtaaact
    12841 cgcaagccga ctgatgcctt ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg
    12901 tctgtaccgg gtgcgttact ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt
    12961 atttccagct acgatcacga caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc
    13021 gatggcgaag gcttcatcgt tattgatgac ctggtggata ccggtggtac tgcggttgcg
    13081 attcgtgaaa tgtatccaaa agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt
    13141 ccgctggttg atgactatgt tgttgatatc ccgcaagata cctggattga acagccgtgg
    13201 gatatgggcg tcgtattcgt cccgccaatc tccggtcgct aatcttttca acgcctggca
    13261 ctgccgggcg ttgttctttt taacttcagg cgggttacaa tagtttccag taagtattct
    13321 ggaggctgca tccatgacac aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa
    13381 acatcctgaa acctcgacgc tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca
    13441 gtcggccctt gatggtaaaa ccatccctca ctggtatcgc atgattaacc gtctgatgtg
    13501 gatctggcgc ggcattgacc cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag
    13561 cgatgccgaa cgtaccgacg atgatttata cgatacggtg attggctacc gtggcggcaa
    13621 ctggatttat gagtgggccc cggatctttg tgaaggaacc ttacttctgt ggtgtgacat
    13681 aattggacaa actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg
    13741 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa
    13801 ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag
    13861 aaatgccatc tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa
    13921 agaagagaaa ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc
    13981 atgctgtgtt tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag
    14041 ctgcactgct atacaagaaa attatggaaa aatattctgt aacctttata agtaggcata
    14101 acagttataa tcataacata ctgttttttc ttactccaca caggcataga gtgtctgcta
    14161 ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt aatttgtaaa ggggttaata
    14221 aggaatattt gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta
    14281 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
    14341 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    14401 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    14461 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
    14521 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
    14581 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
    14641 tgtactacaa acttagtagt
//
