LOCUS Exported 10397 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Contains Golden Gate cassette to express up to four gRNAs and dsRED. ACCESSION . VERSION . KEYWORDS pLV GG hUbC-dsRED SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10397) AUTHORS Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA TITLE Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector. JOURNAL Nucleic Acids Res. 2014 Aug 13. pii: gku749. PUBMED 25122746 REFERENCE 2 (bases 1 to 10397) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10397 /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" protein_bind 2602..2635 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(2622..2948) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATTVSY HNFV" primer_bind 2798..2820 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2812..2829 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2813..2829 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2839..2858 /label=T7 /note="T7 promoter, forward primer" promoter 2839..2857 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2905..2925) /label=T3 /note="T3 promoter, forward primer" promoter complement(2905..2923) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2944..2960) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 2968..2984 /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(2992..3022) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3037..3058 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3124..4334 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 4171..4190 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" regulatory 4381..4390 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4387..5064 /codon_start=1 /product="noncytotoxic tetrameric variant of DsRed fluorescent protein (Strack et al., 2008)" /label=DsRed-Express2 /note="mammalian codon-optimized" /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH LFQ" primer_bind complement(4567..4587) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 4785..4804 /label=mCherry-F /note="mCherry, forward primer" primer_bind 4979..5002 /label=DsRed1-C /note="DsRed1, forward primer" misc_feature 5089..5677 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5142..5162) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5560..5571) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5680..5696) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5681..5697) /label=pBluescriptKS /note="For pBluescript vector" LTR 6202..6382 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(6408..6425) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6414..6638 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6684..7112 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6771..6790) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6981..7002 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7121..7141) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7126..7455 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7306..7441 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7368..7387 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 7503..7550 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 7569..7943 /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 8073..8194 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8110..8129) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8164..8183 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8243..8259) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 8267..8283 /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(8291..8321) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8336..8357 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8474..8491) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8645..9233) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8725..8744) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9404..10264) /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 10027..10046 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10265..10369) /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 aataacttcg tatagcatac attatacgaa gttatgataa 2641 gagacggtgg ttgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt 2701 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 2761 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 2821 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc 2881 ctcgaggtcc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc 2941 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 3001 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 3061 tgcgttgcgc tcactgcccg ctttccactg catgacgtct ccacaattaa ttaagggtgc 3121 agcggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc acggcgagcg 3181 ctgccacgtc agacgaaggg cgcaggagcg ttcctgatcc ttccgcccgg acgctcagga 3241 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt 3301 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg 3361 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc 3421 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg 3481 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagttgc gggctgctgg 3541 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac 3601 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga 3661 gcgcacaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtaa ggcgggctgt 3721 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc 3781 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca tctggggacc 3841 ctgacgtgaa gtttgtcact gactggagaa ctcgggtttg tcgtctggtt gcgggggcgg 3901 cagttatgcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg cctcgtcgtg 3961 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg 4021 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga 4081 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt 4141 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg 4201 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt 4261 caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg 4321 cttttttgtt agacgaagct tgggctgcag gtcgactcta gaggatcccc gggtaccggt 4381 gccaccatgg atagcactga gaacgtcatc aagcccttca tgcgcttcaa ggtgcacatg 4441 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg caagccctac 4501 gagggcaccc agaccgccaa gctgcaggtg accaagggcg gccccctgcc cttcgcctgg 4561 gacatcctgt ccccccagtt ccagtacggc tccaaggtgt acgtgaagca ccccgccgac 4621 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac 4681 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcaccttc 4741 atctaccacg tgaagttcat cggcgtgaac ttcccctccg acggccccgt aatgcagaag 4801 aagactctgg gctgggagcc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag 4861 ggcgagatcc acaaggcgct gaagctgaag ggcggcggcc actacctggt ggagttcaag 4921 tcaatctaca tggccaagaa gcccgtgaag ctgcccggct actactacgt ggactccaag 4981 ctggacatca cctcccacaa cgaggactac accgtggtgg agcagtacga gcgcgccgag 5041 gcccgccacc acctgttcca gtaggaattc gatatcaagc ttatcgataa tcaacctctg 5101 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5161 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5221 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5281 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5341 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5401 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5461 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5521 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5581 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5641 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgagac 5701 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 5761 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 5821 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5881 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 5941 acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca 6001 ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc 6061 aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg 6121 gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca catggcccga 6181 gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc 6241 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa 6301 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 6361 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 6421 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 6481 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 6541 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 6601 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 6661 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 6721 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 6781 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 6841 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 6901 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 6961 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 7021 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 7081 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 7141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 7201 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 7261 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 7321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 7381 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 7441 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac 7501 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga 7561 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg 7621 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact 7681 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg 7741 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt 7801 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg 7861 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact 7921 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct 7981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 8041 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 8101 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 8161 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 8221 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 8281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 8341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 8401 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 8461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 8521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 8581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 8641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 8701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 8761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 8821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 8881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 8941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 9001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 9061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 9121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 9181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 9241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 9301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 9361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 9421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 9481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 9541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 9601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 9661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 9721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 9781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 9841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 9901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 9961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 10021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 10081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 10141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 10201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 10261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 10321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 10381 gaaaagtgcc acctgac //