LOCUS Exported 12990 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CasRx (NLS-RfxCas13d-NLS) with 2A-EGFP for RNA targeting. ACCESSION . VERSION . KEYWORDS pXR001: EF1a-CasRx-2A-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12990) AUTHORS Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD TITLE Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. JOURNAL Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. PUBMED 29551272 REFERENCE 2 (bases 1 to 12990) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12990 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /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 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2682..3859 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2913..3850 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3807..3827 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3878..3898 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3908..6805 /codon_start=1 /product="RNA-targeting Type VI CRISPR protein from Ruminococcus flavefaciens strain XPD3002 (Konermann et al., 2018)" /label=RfxCas13d /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS" CDS 6815..6835 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6845..6871 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 6887..6940 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 6941..7657 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 6941..7654 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(6983..7004) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7244..7263) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7591..7612 /label=EGFP-C /note="EGFP, forward primer" misc_feature 7682..8270 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7735..7755) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8153..8164) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8273..8289) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8274..8290) /label=pBluescriptKS /note="For pBluescript vector" LTR 8795..8975 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(9001..9018) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9007..9231 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 9277..9705 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9364..9383) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9574..9595 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9714..9734) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9719..10048 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9899..10034 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9961..9980 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 10096..10143 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 10162..10536 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 10666..10787 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10703..10722) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10757..10776 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10836..10852) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10836..10852) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10849..10871) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10860..10876 /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(10884..10914) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10929..10950 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11067..11084) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11238..11826) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11318..11337) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11997..12857) /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 12620..12639 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12858..12962) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt 2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt 2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact ggctgcagta 3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc 3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg 3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat 3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc 3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg 3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg 3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat 3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg 3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg 3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg 3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag 3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg 3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct 3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct 3841 tccatttcag gtgtcgtgac gtacggccac catgagcccc aagaagaaga gaaaggtgga 3901 ggccagcatc gaaaaaaaaa agtccttcgc caagggcatg ggcgtgaagt ccacactcgt 3961 gtccggctcc aaagtgtaca tgacaacctt cgccgaaggc agcgacgcca ggctggaaaa 4021 gatcgtggag ggcgacagca tcaggagcgt gaatgagggc gaggccttca gcgctgaaat 4081 ggccgataaa aacgccggct ataagatcgg caacgccaaa ttcagccatc ctaagggcta 4141 cgccgtggtg gctaacaacc ctctgtatac aggacccgtc cagcaggata tgctcggcct 4201 gaaggaaact ctggaaaaga ggtacttcgg cgagagcgct gatggcaatg acaatatttg 4261 tatccaggtg atccataaca tcctggacat tgaaaaaatc ctcgccgaat acattaccaa 4321 cgccgcctac gccgtcaaca atatctccgg cctggataag gacattattg gattcggcaa 4381 gttctccaca gtgtatacct acgacgaatt caaagacccc gagcaccata gggccgcttt 4441 caacaataac gataagctca tcaacgccat caaggcccag tatgacgagt tcgacaactt 4501 cctcgataac cccagactcg gctatttcgg ccaggccttt ttcagcaagg agggcagaaa 4561 ttacatcatc aattacggca acgaatgcta tgacattctg gccctcctga gcggactgag 4621 gcactgggtg gtccataaca acgaagaaga gtccaggatc tccaggacct ggctctacaa 4681 cctcgataag aacctcgaca acgaatacat ctccaccctc aactacctct acgacaggat 4741 caccaatgag ctgaccaact ccttctccaa gaactccgcc gccaacgtga actatattgc 4801 cgaaactctg ggaatcaacc ctgccgaatt cgccgaacaa tatttcagat tcagcattat 4861 gaaagagcag aaaaacctcg gattcaatat caccaagctc agggaagtga tgctggacag 4921 gaaggatatg tccgagatca ggaaaaatca taaggtgttc gactccatca ggaccaaggt 4981 ctacaccatg atggactttg tgatttatag gtattacatc gaagaggatg ccaaggtggc 5041 tgccgccaat aagtccctcc ccgataatga gaagtccctg agcgagaagg atatctttgt 5101 gattaacctg aggggctcct tcaacgacga ccagaaggat gccctctact acgatgaagc 5161 taatagaatt tggagaaagc tcgaaaatat catgcacaac atcaaggaat ttaggggaaa 5221 caagacaaga gagtataaga agaaggacgc ccctagactg cccagaatcc tgcccgctgg 5281 ccgtgatgtt tccgccttca gcaaactcat gtatgccctg accatgttcc tggatggcaa 5341 ggagatcaac gacctcctga ccaccctgat taataaattc gataacatcc agagcttcct 5401 gaaggtgatg cctctcatcg gagtcaacgc taagttcgtg gaggaatacg cctttttcaa 5461 agactccgcc aagatcgccg atgagctgag gctgatcaag tccttcgcta gaatgggaga 5521 acctattgcc gatgccagga gggccatgta tatcgacgcc atccgtattt taggaaccaa 5581 cctgtcctat gatgagctca aggccctcgc cgacaccttt tccctggacg agaacggaaa 5641 caagctcaag aaaggcaagc acggcatgag aaatttcatt attaataacg tgatcagcaa 5701 taaaaggttc cactacctga tcagatacgg tgatcctgcc cacctccatg agatcgccaa 5761 aaacgaggcc gtggtgaagt tcgtgctcgg caggatcgct gacatccaga aaaaacaggg 5821 ccagaacggc aagaaccaga tcgacaggta ctacgaaact tgtatcggaa aggataaggg 5881 caagagcgtg agcgaaaagg tggacgctct cacaaagatc atcaccggaa tgaactacga 5941 ccaattcgac aagaaaagga gcgtcattga ggacaccggc agggaaaacg ccgagaggga 6001 gaagtttaaa aagatcatca gcctgtacct caccgtgatc taccacatcc tcaagaatat 6061 tgtcaatatc aacgccaggt acgtcatcgg attccattgc gtcgagcgtg atgctcaact 6121 gtacaaggag aaaggctacg acatcaatct caagaaactg gaagagaagg gattcagctc 6181 cgtcaccaag ctctgcgctg gcattgatga aactgccccc gataagagaa aggacgtgga 6241 aaaggagatg gctgaaagag ccaaggagag cattgacagc ctcgagagcg ccaaccccaa 6301 gctgtatgcc aattacatca aatacagcga cgagaagaaa gccgaggagt tcaccaggca 6361 gattaacagg gagaaggcca aaaccgccct gaacgcctac ctgaggaaca ccaagtggaa 6421 tgtgatcatc agggaggacc tcctgagaat tgacaacaag acatgtaccc tgttcagaaa 6481 caaggccgtc cacctggaag tggccaggta tgtccacgcc tatatcaacg acattgccga 6541 ggtcaattcc tacttccaac tgtaccatta catcatgcag agaattatca tgaatgagag 6601 gtacgagaaa agcagcggaa aggtgtccga gtacttcgac gctgtgaatg acgagaagaa 6661 gtacaacgat aggctcctga aactgctgtg tgtgcctttc ggctactgta tccccaggtt 6721 taagaacctg agcatcgagg ccctgttcga taggaacgag gccgccaagt tcgacaagga 6781 gaaaaagaag gtgtccggca attccggatc cggacctaag aaaaagagga aggtggcggc 6841 cgcttaccca tacgatgttc cagattacgc tgctagcggc agtggagagg gcagaggaag 6901 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca gtgagcaagg gcgaggagct 6961 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt 7021 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat 7081 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg 7141 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc 7201 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa 7261 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg 7321 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag 7381 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat 7441 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc 7501 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct 7561 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 7621 cgggatcact ctcggcatgg acgagctgta caagtaagaa ttcgatatca agcttatcga 7681 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 7741 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 7801 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 7861 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 7921 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 7981 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 8041 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 8101 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 8161 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 8221 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 8281 tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa 8341 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc 8401 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa 8461 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct 8521 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg 8581 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa 8641 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg 8701 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca 8761 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg 8821 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 8881 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 8941 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag 9001 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 9061 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 9121 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 9181 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 9241 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 9301 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 9361 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 9421 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 9481 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 9541 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 9601 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 9661 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 9721 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 9781 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 9841 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 9901 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 9961 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 10021 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 10081 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac 10141 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg 10201 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc 10261 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc 10321 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag 10381 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc 10441 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc 10501 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc 10561 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 10621 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 10681 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 10741 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 10801 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 10861 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 10921 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 10981 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 11041 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 11101 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 11161 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 11221 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 11281 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 11341 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 11401 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 11461 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 11521 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 11581 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 11641 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 11701 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 11761 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 11821 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 11881 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 11941 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 12001 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 12061 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 12121 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 12181 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 12241 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 12301 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 12361 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 12421 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 12481 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 12541 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 12601 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 12661 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 12721 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 12781 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 12841 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 12901 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 12961 cgcacatttc cccgaaaagt gccacctgac //