LOCUS Exported 10947 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION lenti vector encoding the MS2-VP64 with a 2A Hygro resistance marker. ACCESSION . VERSION . KEYWORDS Lenti_MCP-VP64_Hygro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10947) AUTHORS Li K, Liu Y, Cao H, Zhang Y, Gu Z, Liu X, Yu A, Kaphle P, Dickerson KE, Ni M, Xu J TITLE Interrogation of enhancer function by enhancer-targeting CRISPR epigenetic editing. JOURNAL Nat Commun. 2020 Jan 24;11(1):485. doi: 10.1038/s41467-020-14362-5. PUBMED 31980609 REFERENCE 2 (bases 1 to 10947) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10947 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..580 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 598..778 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 825..950 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1443..1676 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1861..1905 /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 2203..2320 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2450..3628 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2681..3619 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3576..3596 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3635..3644 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3644..4030 /codon_start=1 /product="bacteriophage MS2 coat protein" /label=MS2-N55K /note="binds with high affinity to a specific stem-loop structure in the viral RNA (Lim et al., 1994)" /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI VKAMQGLLKDGNPIPSAIAANSGIY" CDS 4085..4105 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4121..4270 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 4286..4339 /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 4343..5362 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK " misc_feature 5402..5990 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5455..5475) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5873..5884) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5993..6009) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5994..6010) /label=pBluescriptKS /note="For pBluescript vector" LTR 6515..6695 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(6721..6738) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6727..6951 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6997..7425 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7084..7103) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7294..7315 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7434..7454) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7439..7768 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7619..7754 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7681..7700 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 7816..7863 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 7882..8256 /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 8386..8507 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8423..8442) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8477..8496 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8556..8572) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8556..8572) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8569..8591) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8580..8596 /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(8604..8634) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8649..8670 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8787..8804) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8958..9546) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9038..9057) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9717..10577) /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 10340..10359 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10578..10682) /gene="bla" /label=AmpR promoter primer_bind complement(10757..10776) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg 601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga 841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa 901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc 961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat 1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg 1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc 1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc 1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa 1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat 1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta 1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga 1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca 1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat 1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag 1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg 1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg 1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa 1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa 1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac 1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt 1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca 2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa 2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg 2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg 2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac 2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag 2341 cagagatcca gtttggttaa ttagctagct gcaaagatgg ataaagtttt aaacagagag 2401 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc 2461 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg 2521 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta 2581 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt 2641 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc 2701 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg 2761 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag 2821 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg 2881 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc 2941 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct 3001 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga 3061 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc 3121 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc 3181 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc 3241 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatgaagga cgcggcgctc 3301 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt 3361 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag 3421 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca 3481 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga 3541 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag 3601 tttttttctt ccatttcagg tgtcgtgacg tacggccacc atggcttcaa actttactca 3661 gttcgtgctc gtggacaatg gtgggacagg ggatgtgaca gtggctcctt ctaatttcgc 3721 taatggggtg gcagagtgga tcagctccaa ctcacggagc caggcctaca aggtgacatg 3781 cagcgtcagg cagtctagtg cccagaagag aaagtatacc atcaaggtgg aggtccccaa 3841 agtggctacc cagacagtgg gcggagtcga actgcctgtc gccgcttgga ggtcctacct 3901 gaacatggag ctcactatcc caattttcgc taccaattct gactgtgaac tcatcgtgaa 3961 ggcaatgcag gggctcctca aagacggtaa tcctatccct tccgccatcg ccgctaactc 4021 aggtatctac agcgctggag gaggtggaag cggaggagga ggaagcggag gaggaggtag 4081 cggacctaag aaaaagagga aggtggcggc cgctggatcc gatgctctcg acgattttga 4141 cctcgatatg ctgggatccg acgctctcga tgacttcgac cttgacatgc tggggagtga 4201 cgctctggac gacttcgatc tcgacatgct gggctccgac gccctggatg acttcgatct 4261 ggacatgctc tgtacaggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt 4321 cgaggagaat cctggcccaa ccatgaaaaa gcctgaactc accgctacct ctgtcgagaa 4381 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctccg agggcgaaga 4441 atctcgggct ttcagcttcg atgtgggagg gcgtggatat gtcctgcggg tgaatagctg 4501 cgccgatggt ttctacaaag atcgctatgt ttatcggcac tttgcatccg ccgctctccc 4561 tattcccgaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg 4621 ccgtgcacag ggtgtcacct tgcaagacct gcctgaaacc gaactgcccg ctgttctcca 4681 gcccgtcgcc gaggccatgg atgccatcgc tgccgccgat cttagccaga ccagcgggtt 4741 cggcccattc ggacctcaag gaatcggtca atacactaca tggcgcgatt tcatctgcgc 4801 tattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcctc 4861 cgtcgcccag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca 4921 cctcgtgcac gccgatttcg gctccaacaa tgtcctgacc gacaatggcc gcataacagc 4981 cgtcattgac tggagcgagg ccatgttcgg ggattcccaa tacgaggtcg ccaacatctt 5041 cttctggagg ccctggttgg cttgtatgga gcagcagacc cgctacttcg agcggaggca 5101 tcccgagctt gcaggatctc ctcggctccg ggcttatatg ctccgcattg gtcttgacca 5161 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggctc agggtcgctg 5221 cgacgcaatc gtccggtccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag 5281 cgctgccgtc tggaccgatg gctgtgtgga agtgctcgcc gatagtggaa acagacgccc 5341 cagcactcgt cctagggcaa aggatctgca gtaatgagaa ttcgatatca agcttatcgg 5401 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5461 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5521 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5581 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5641 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5701 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5761 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5821 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5881 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5941 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 6001 tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa 6061 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc 6121 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa 6181 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct 6241 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg 6301 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa 6361 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg 6421 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca 6481 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg 6541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 6601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 6661 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag 6721 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 6781 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 6841 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 6901 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 6961 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 7021 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 7081 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 7141 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 7201 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 7261 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 7321 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 7381 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 7441 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 7501 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 7561 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 7621 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 7681 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 7741 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 7801 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac 7861 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg 7921 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc 7981 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc 8041 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag 8101 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc 8161 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc 8221 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc 8281 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 8341 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 8401 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 8461 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 8521 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 8581 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 8641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 8701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 8761 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 8821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 8881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 8941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 9001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 9061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 9121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 9181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 9241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 9301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 9361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 9421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 9481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 9541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 9601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 9661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 9721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 9781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 9841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 9901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 9961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 10021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 10081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 10141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 10201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 10261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 10321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 10381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 10441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 10501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 10561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 10621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 10681 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc 10741 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc 10801 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc 10861 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc 10921 gcgatgtacg ggccagatat acgcgtt //