LOCUS Exported 9603 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 9603) 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 9603) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9603 /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..2851 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 2770..2788 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 2861..3652 /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(2915..2934) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3525..3544 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS 3662..3715 /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 3716..4462 /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(4502..4522) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4920..4931) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5136..5369 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5441..5562 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5478..5497) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5532..5551 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5602..5737 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5664..5683 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 5737..5808 /gene="bla" /label=AmpR promoter CDS 5809..6669 /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(6027..6046) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6840..7428 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7329..7348 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7582..7599 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7716..7737 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7752..7782 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7790..7806 /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 7795..7817 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7814..7830 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7814..7830 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7849..7869 /label=T3 /note="T3 promoter, forward primer" promoter 7851..7869 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7897..8123 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 8124..8304 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 8351..8476 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8969..9202 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 9387..9431 /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 tctctcccca gcaattcacc atgattgaac aagatggatt 2881 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 2941 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 3001 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 3061 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 3121 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 3181 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 3241 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 3301 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 3361 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac 3421 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 3481 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 3541 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 3601 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tccccggtgc 3661 cgaaggtaga ggttctctcc tcacttgtgg tgatgttgaa gaaaaccctg gtccaatgtc 3721 tagactggac aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat 3781 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta 3841 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag 3901 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa 3961 cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg 4021 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca 4081 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact 4141 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac 4201 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc 4261 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa 4321 gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc 4381 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga 4441 ccttgacatg ctccccgggt aactaagtaa ggatcgatcc aagatatcgt attcttaact 4501 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 4561 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 4621 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 4681 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 4741 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 4801 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 4861 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 4921 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 4981 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 5041 tttcgcctcg gcgtccggac tagaggtacc tttaagacca atgacttaca aggcagctgt 5101 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 5161 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 5221 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 5281 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 5341 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 5401 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 5461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 5581 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 5641 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 5701 ctattccaga agtagtgagg aggctttttt ggaggccttt caaatatgta tccgctcatg 5761 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 5821 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5881 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 5941 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 6001 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 6061 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 6121 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 6181 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 6241 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 6301 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 6361 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 6421 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 6481 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 6541 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 6601 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 6661 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 6721 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 6781 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 6841 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 6901 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 6961 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 7021 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 7081 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 7141 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 7201 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 7261 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 7321 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 7381 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 7441 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 7501 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 7561 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 7621 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 7681 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag 7741 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga 7801 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct 7861 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt 7921 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc 7981 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga 8041 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta 8101 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc 8161 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 8221 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 8281 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa 8341 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga 8401 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga 8461 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg 8521 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga 8581 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat 8641 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa 8701 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc 8761 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc 8821 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat 8881 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg 8941 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag 9001 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt 9061 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt 9121 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat 9181 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca 9241 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca 9301 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa 9361 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat 9421 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca 9481 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga 9541 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg 9601 gac //