LOCUS Exported 11555 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A lentiviral dual fluorescent protein reporter, HypoxCR, detects hypoxic [hypoxia-inducible factor (HIF) active] and/or cycling cells.. ACCESSION . VERSION . KEYWORDS HypoxCR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11555) AUTHORS Le A, Stine ZE, Nguyen C, Afzal J, Sun P, Hamaker M, Siegel NM, Gouw AM, Kang BH, Yu SH, Cochran RL, Sailor KA, Song H, Dang CV TITLE Tumorigenicity of hypoxic respiring cancer cells revealed by a hypoxia-cell cycle dual reporter. JOURNAL Proc Natl Acad Sci U S A. 2014 Aug 11. pii: 201402012. PUBMED 25114222 REFERENCE 2 (bases 1 to 11555) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11555 /organism="synthetic DNA construct" /mol_type="other DNA" 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 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" polyA_signal 2240..2464 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 2453..2470 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" CDS complement(2486..3334) /codon_start=1 /product="EGFP destabilized by fusion to a mutated version of residues 422-461 of mouse ornithine decarboxylase, giving an in vivo half-life of ~1 hour" /label=d1EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV I" primer_bind complement(2660..2681) /label=EGFP-C /note="EGFP, forward primer" primer_bind 3009..3028 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3268..3289 /label=EGFP-N /note="EGFP, reverse primer" regulatory 3331..3340 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(3373..3411) /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" enhancer 4636..4939 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4940..5143 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5094..5114 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5272..5281 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5278..5985 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(5423..5441) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 5691..5710 /label=mCherry-F /note="mCherry, forward primer" CDS 6016..6345 /codon_start=1 /product="degron consisting of residues 1-110 of human Geminin (Zielke and Edgar, 2015)" /label=Gem1 (1-110) /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE KRRKAL" CDS 6346..6369 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter 6389..6888 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(6424..6447) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 6821..6840 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6909..7508 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6909..6928) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 7405..7425 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 7522..8110 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7575..7595) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7993..8004) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8317..8950 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(9079..9095) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9079..9095) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9092..9114) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9103..9119 /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(9127..9157) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9172..9193 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(9310..9327) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9481..10069) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9561..9580) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10240..11100) /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 10863..10882 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(11101..11205) /gene="bla" /label=AmpR promoter polyA_signal 11253..11387 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11290..11309) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 11344..11363 /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 tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatcagggc gcgtggggat accccctaga gccccagctg 2221 gttctttccg cctcagaagc catagagccc gggccatccc cagcatgcct gctattgtct 2281 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta 2341 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct 2401 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc 2461 acagtcgagg ctgatcagcg agcttctaga tcacattgat cctagcagaa gcacaggctg 2521 cagggtgacg gtccatcccg ctctcctggg cacaagacat gggcagcgtg ccatcatcct 2581 gcgccgccac cgccggcggg aagccatggc taagcttctt gtacagctcg tccatgccga 2641 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg 2701 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc 2761 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga 2821 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat 2881 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct 2941 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct 3001 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc 3061 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga 3121 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc 3181 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc 3241 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca 3301 ccccggtgaa cagctcctcg cccttgctca ccatggtggc aagcttctag tgatctgacg 3361 gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac cagatctcga 3421 caagaggacc tgttggagcc cacgtatgca ctgtggctcg acaagaggac ctgttggagc 3481 ccacgtatgc actgtggctc gagaaaagct cggtaccgat ctcttaatta agagctcacg 3541 gggacagccc gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct 3601 gctctttgag cctgcagaca cctgggggga tacggggaaa aagctttagg ctgaaagaga 3661 gatttagaat gacagaatca tagaacggcc tgggttgcaa aggagcacag tgctcatcca 3721 gatccaaccc cctgctatgt gcagggtcat caaccagcag cccaggctgc ccagagccac 3781 atccagcctg gccttgaatg cctgcaggga tggggcatcc acagcctcct tgggcaacct 3841 gttcagtgcg tcaccaccct ctgggggaaa aactgcctcc tcatatccaa cccaaacctc 3901 ccctgtctca gtgtaaagcc attccccctt gtcctatcaa gggggagttt gctgtgacat 3961 tgttggtctg gggtgacaca tgtttgccaa ttcagtgcat cacggagagg cagatcttgg 4021 ggataaggaa gtgcaggaca gcatggacgt gggacatgca ggtgttgagg gctctgggac 4081 actctccaag tcacagcgtt cagaacagcc ttaaggataa gaagatagga tagaaggaca 4141 aagagcaagt taaaacccag catggagagg agcacaaaaa ggccacagac actgctggtc 4201 cctgtgtctg agcctgcatg tttgatggtg tctggatgca agcagaaggg gtggaagagc 4261 ttgcctggag agatacagct gggtcagtag gactgggaca ggcagctgga gaattgccat 4321 gtagatgttc atacaatcgt caaatcatga aggctggaaa agccctccaa gatccccaag 4381 accaacccca acccacccac cgtgcccact ggccatgtcc ctcagtgcca catccccaca 4441 gttcttcatc acctccaggg acggtgaccc ccccacctcc gtgggcagct gtgccactgc 4501 agcaccgctc tttggagaag gtaaatcttg ctaaatccag cccgaccctc ccctggcaca 4561 acgtaaggcc attatctctc atccaactcc aggacggagt cagtgagaat attatcgata 4621 agcttgggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 4681 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 4741 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 4801 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 4861 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 4921 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 4981 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 5041 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 5101 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca 5161 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gactctacta 5221 gaggatcgct agcgctaccg gactcagatc tcgagctcaa gcttcgaatt cgccaccatg 5281 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg 5341 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc 5401 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc 5461 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc 5521 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg 5581 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc 5641 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg 5701 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc 5761 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag 5821 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac 5881 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc 5941 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagggata tccatcacac 6001 tggcggccgc tcgagatgaa cccatccatg aagcaaaagc aggaggagat taaggaaaac 6061 atcaaaaact ccagtgtgcc tcgccggacc ctcaaaatga tccaaccatc cgccagcggc 6121 agcctcgtgg gtcgcgagaa cgaattgagc gccggactga gcaagcgcaa gcacagaaac 6181 gatcatctca ccagcactac aagctctcca ggcgtgatcg tgccggagag ctccgagaac 6241 aagaacttgg gcggtgtgac acaagaaagc ttcgacctga tgatcaagga gaaccctagt 6301 agccaatact ggaaagaggt cgccgagaag agacggaaag ccctcgacta caaagacgat 6361 gacgacaagt agtctagata attctaccgg gtaggggagg cgcttttccc aaggcagtct 6421 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct 6481 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc 6541 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg 6601 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc 6661 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc 6721 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc 6781 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag 6841 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca 6901 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca 6961 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 7021 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 7081 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 7141 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 7201 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 7261 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 7321 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 7381 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 7441 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 7501 gtgcctgacc gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact 7561 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 7621 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 7681 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 7741 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 7801 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 7861 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 7921 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 7981 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 8041 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 8101 gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa 8161 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg 8221 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg 8281 tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac 8341 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc 8401 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc 8461 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt 8521 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt 8581 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga 8641 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg 8701 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt 8761 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta 8821 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc 8881 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa 8941 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa 9001 tgaatatcag agagtgagag gccttgacat tgctagcgtt ttaccgtcga cctctagcta 9061 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 9121 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 9181 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 9241 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 9301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 9361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 9421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 9481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 9541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 9601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 9661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 9721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 9781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 9841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 9901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 9961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 10021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 10081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 10141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 10201 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 10261 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 10321 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 10381 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 10441 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 10501 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 10561 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 10621 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 10681 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 10741 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 10801 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 10861 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 10921 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 10981 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 11041 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 11101 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 11161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 11221 agtgccacct gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt 11281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 11341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc 11401 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc 11461 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt 11521 gtatttgtta aatatgtact acaaacttag tagtt //