LOCUS Exported 9602 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tet-On construct to induce expression of tandem eGFP-mCherry tagged RAMP4 (ER marker). ACCESSION . VERSION . KEYWORDS TetOn-mCherry-eGFP-RAMP4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9602) AUTHORS Liang JR, Lingeman E, Ahmed S, Corn JE TITLE Atlastins remodel the endoplasmic reticulum for selective autophagy. JOURNAL J Cell Biol. 2018 Aug 24. pii: jcb.201804185. doi: 10.1083/jcb.201804185. PUBMED 30143524 REFERENCE 2 (bases 1 to 9602) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9602 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 41..158 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 212..526 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 220..238 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 256..274 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 291..309 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 327..345 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 363..381 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 398..416 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 434..452 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 499..523 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 586..595 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 592..1299 /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(737..755) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1005..1024 /label=mCherry-F /note="mCherry, forward primer" protein_bind 1327..1351 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 1360..1369 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1366..2082 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1366..2082 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1411..1432) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1672..1691) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2019..2040 /label=EGFP-C /note="EGFP, forward primer" promoter 2341..2824 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 2770..2788 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 2860..3651 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2914..2933) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3524..3543 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS 3661..3714 /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 3715..4461 /codon_start=1 /product="improved tetracycline-controlled transactivator" /label=rtTA-Advanced /note="In the Tet-On(R) system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline." /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" primer_bind complement(4501..4521) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4919..4930) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5135..5368 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5440..5561 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5477..5496) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5531..5550 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5601..5736 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5663..5682 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 5736..5807 /gene="bla" /label=AmpR promoter CDS 5808..6668 /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 complement(6026..6045) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6839..7427 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7328..7347 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7581..7598 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7715..7736 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7751..7781 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7789..7805 /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)." primer_bind 7794..7816 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7813..7829 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7813..7829 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7848..7868 /label=T3 /note="T3 promoter, forward primer" promoter 7850..7868 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7896..8122 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 8123..8303 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 8350..8475 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8968..9201 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 9386..9430 /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" ORIGIN 1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga 61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta 121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca 181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac 241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag 301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt 361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt 421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg 481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta 541 gcgccacccc agccctcact ccttctctag gcgccggccg gatctgccac catggtgagc 601 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg 661 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac 721 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg 781 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac 841 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac 901 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc 961 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag 1021 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag 1081 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag 1141 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag 1201 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag 1261 ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggactcag atccgctggc 1321 ccaggaacaa gtttgtacaa aaaagcaggc tttccggtcg ccaccatggt gagcaagggc 1381 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 1441 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 1501 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 1561 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc 1621 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 1681 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 1741 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 1801 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac 1861 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag 1921 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 1981 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 2041 accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact cagatccggt 2101 accgaattcg gtggatctgg aggttctggt ggaatggtcg ccaagcaaag gatccgtatg 2161 gccaacgaga agcacagcaa gaacatcacc cagcgcggca acgtcgccaa gacctcgaga 2221 aatgcccccg aagagaaggc gtctgtagga ccctggttat tggctctctt catttttgtt 2281 gtctgtggtt ctgcaatttt ccagattatt caaagtatca ggatgggcat gtgaggatcc 2341 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc 2401 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc 2461 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc 2521 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac 2581 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc 2641 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag 2701 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct 2761 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct 2821 cgttgaccga atcaccgacc tcttccccag caattcacca tgattgaaca agatggattg 2881 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 2941 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 3001 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 3061 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3121 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3181 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3241 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3301 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 3361 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 3421 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 3481 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 3541 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 3601 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ccccggtgcc 3661 gaaggtagag gttctctcct cacttgtggt gatgttgaag aaaaccctgg tccaatgtct 3721 agactggaca agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc 3781 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac 3841 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg 3901 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac 3961 gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc 4021 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag 4081 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg 4141 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc 4201 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc 4261 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag 4321 tgcgaaagcg gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc 4381 gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac 4441 cttgacatgc tccccgggta actaagtaag gatcgatcca agatatcgta ttcttaacta 4501 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 4561 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 4621 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 4681 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 4741 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 4801 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 4861 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 4921 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 4981 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgt 5041 ttcgcctcgg cgtccggact agaggtacct ttaagaccaa tgacttacaa ggcagctgta 5101 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga 5161 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg 5221 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 5281 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 5341 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 5401 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat 5461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 5521 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta 5581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 5641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 5701 tattccagaa gtagtgagga ggcttttttg gaggcctttc aaatatgtat ccgctcatga 5761 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 5821 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 5881 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 5941 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 6001 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 6061 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 6121 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 6181 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 6241 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 6301 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 6361 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 6421 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 6481 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 6541 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 6601 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 6661 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 6721 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 6781 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 6841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 6901 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 6961 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 7021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 7081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 7141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 7201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 7261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 7321 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 7381 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 7441 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 7501 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 7561 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 7621 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 7681 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg 7741 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat 7801 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc 7861 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta 7921 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg 7981 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac 8041 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa 8101 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct 8161 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 8221 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 8281 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa 8341 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag 8401 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat 8461 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg 8521 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag 8581 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata 8641 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat 8701 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct 8761 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct 8821 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata 8881 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt 8941 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc 9001 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc 9061 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt 9121 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata 9181 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac 9241 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac 9301 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat 9361 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg 9421 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat 9481 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa 9541 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg 9601 ac //