LOCUS Exported 10415 bp ds-DNA circular SYN 08-JUN-2021 DEFINITION Barcoded lentiviral vector to express CDK4 (R24C) in mammalian cells under control of EF1a promoter. Barcoded for transcript detection in single cell RNA-seq. ACCESSION . VERSION . KEYWORDS LV_EF1a_CDK4(R24C)-P2A-Hygro_Barcode SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10415) TITLE Mali Lab Plasmids REFERENCE 2 (bases 1 to 10415) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 8, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10415 /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 2693..3871 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2924..3862 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3819..3839 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 3888..3907 /label=T7 /note="T7 promoter, forward primer" promoter 3888..3906 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 3964..3973 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4888..4944 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 4945..5970 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" protein_bind 6162..6195 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." LTR 6220..6400 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(6426..6443) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6432..6656 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6702..7130 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6789..6808) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6999..7020 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7139..7159) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7144..7473 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7324..7459 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7386..7405 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 7521..7568 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 7587..7961 /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 8091..8212 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8128..8147) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8182..8201 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8261..8277) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8261..8277) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8274..8296) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8285..8301 /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(8309..8339) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8354..8375 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8492..8509) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8663..9251) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8743..8762) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9422..10282) /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 10045..10064 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10283..10387) /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 atcgataagc tttgcaaaga tggataaagt tttaaacaga 2641 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg 2701 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 2761 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 2821 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 2881 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg 2941 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac 3001 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc 3061 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg 3121 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa 3181 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag 3241 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg 3301 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc 3361 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc 3421 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg 3481 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg 3541 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc 3601 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc 3661 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc 3721 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt 3781 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca 3841 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac 3901 tatagggaga cccaagctgg ctaggtaagc ttgatatcga attcctgcag cccgggggat 3961 cccgccacca tggctacctc tcgatatgag ccagtggctg aaattggtgt cggtgcctat 4021 gggacagtgt acaaggcctg tgatccccac agtggccact ttgtggccct caagagtgtg 4081 agagtcccca atggaggagg aggtggagga ggccttccca tcagcacagt tcgtgaggtg 4141 gctttactga ggcgactgga ggcttttgag catcccaatg ttgtccggct gatggacgtc 4201 tgtgccacat cccgaactga ccgggagatc aaggtaaccc tggtgtttga gcatgtagac 4261 caggacctaa ggacatatct ggacaaggca cccccaccag gcttgccagc cgaaacgatc 4321 aaggatctga tgcgccagtt tctaagaggc ctagatttcc ttcatgccaa ttgcatcgtt 4381 caccgagatc tgaagccaga gaacattctg gtgacaagtg gtggaacagt caagctggct 4441 gactttggcc tggccagaat ctacagctac cagatggcac ttacacccgt ggttgttaca 4501 ctctggtacc gagctcccga agttcttctg cagtccacat atgcaacacc tgtggacatg 4561 tggagtgttg gctgtatctt tgcagagatg tttcgtcgaa agcctctctt ctgtggaaac 4621 tctgaagccg accagttggg caaaatcttt gacctgattg ggctgcctcc agaggatgac 4681 tggcctcgag atgtatccct gccccgtgga gcctttcccc ccagagggcc ccgcccagtg 4741 cagtcggtgg tacctgagat ggaggagtcg ggagcacagc tgctgctgga aatgctgact 4801 tttaacccac acaagcgaat ctctgccttt cgagctctgc agcactctta tctacataag 4861 gatgaaggta atccggaggg atccggcgca acaaacttct ctctgctgaa acaagccgga 4921 gatgtcgaag agaatcctgg accgatgaaa aagcctgaac tcaccgcgac gtctgtcgag 4981 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa 5041 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc 5101 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc 5161 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc 5221 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg 5281 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg 5341 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc 5401 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg 5461 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg 5521 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca 5581 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc 5641 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg 5701 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac 5761 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga 5821 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga 5881 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc 5941 cccagcactc gtccgagggc aaaggaataa gttagaacta tttcctggct gttacgcgtg 6001 attgtcaata gcgttattct actgacgggc ttgttaaaca gtgaaccgcc cgccccacga 6061 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgaacaaaa gaaaaggggg 6121 gactggaagg gctaattcac tcccaacgaa gacaagatat cataacttcg tatagcatac 6181 attatacgaa gttatcggct agctggtccg gactgtactg ggtctctctg gttagaccag 6241 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 6301 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 6361 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg 6421 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 6481 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 6541 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 6601 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 6661 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 6721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 6781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 6841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 6901 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 6961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 7021 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 7081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 7141 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 7201 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 7261 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 7321 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 7381 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 7441 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 7501 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat 7561 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca 7621 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg 7681 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg 7741 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg 7801 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc 7861 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg 7921 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg 7981 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 8041 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 8101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 8161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 8221 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 8281 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 8341 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 8401 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 8461 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 8521 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 8581 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 8641 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 8701 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 8761 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 8821 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 8881 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 8941 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 9001 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 9061 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 9121 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 9181 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 9241 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 9301 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 9361 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 9421 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 9481 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 9541 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 9601 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 9661 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 9721 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 9781 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 9841 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 9901 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 9961 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 10021 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 10081 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 10141 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 10201 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 10261 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 10321 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 10381 ttccgcgcac atttccccga aaagtgccac ctgac //