LOCUS Exported 12597 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION promoter-inducible expression of constitutively-active RhoA. ACCESSION . VERSION . KEYWORDS pSLIK CA RhoA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12597) AUTHORS MacKay JL, Kumar S TITLE Simultaneous and independent tuning of RhoA and Rac1 activity with orthogonally inducible promoters. JOURNAL Integr Biol (Camb). 2014 Sep;6(9):885-94. doi: 10.1039/c4ib00099d. PUBMED 25044255 REFERENCE 2 (bases 1 to 12597) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12597 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(34..53) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 225..604 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 606..804 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 759..779 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 822..1002 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1049..1174 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1667..1900 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2085..2129 /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 2427..2544 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2601..2625 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter 2657..2971 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 2665..2683 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2701..2719 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2736..2754 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2772..2790 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2808..2826 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2843..2861 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2879..2897 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 2944..2968 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2998..3027 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3034..3615 /codon_start=1 /product="constitutively active mutant of human Rho family small GTPase RhoA" /label=RhoA (Q63L) /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT KDGVREVFEMATRAALQARRGKKKSGCLVL" protein_bind complement(3694..3718) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter 3768..4979 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 4816..4835 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 5104..5850 /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" misc_feature 5903..6476 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(6070..6087) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 6297..6316 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 6504..7223 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6549..6570) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6810..6829) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7157..7178 /label=EGFP-C /note="EGFP, forward primer" misc_feature 7272..7860 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7325..7345) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7743..7754) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(7863..7879) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7864..7880) /label=pBluescriptKS /note="For pBluescript vector" LTR 8389..8569 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(8595..8612) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8601..8825 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 8871..9299 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8958..8977) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9168..9189 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9308..9328) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9313..9642 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9493..9628 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9555..9574 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 9690..9737 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 9756..10130 /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 10260..10381 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10297..10316) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10351..10370 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10430..10446) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10430..10446) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10443..10465) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10454..10470 /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(10478..10508) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10523..10544 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10661..10678) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10832..11420) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10912..10931) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11591..12451) /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 12214..12233 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12452..12556) /gene="bla" /label=AmpR promoter ORIGIN 1 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt 61 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat 121 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag 181 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac 241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 781 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc 841 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 901 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 961 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga 1021 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc 1081 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg 1141 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc 1201 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat 1261 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc 1321 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga 1381 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat 1441 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac 1501 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga 1561 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca 1621 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc 1681 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac 1741 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg 1801 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa 1861 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg 1921 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg 1981 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa 2041 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat 2101 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt 2161 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg 2221 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc 2281 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga 2341 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc 2401 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt 2461 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa 2521 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg 2581 ttaattctag acagttatca acaagtttgt acaaaaaagc aggctttaaa ggaaccaatt 2641 cagtcgactg gatcccgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac 2701 tccctatcag tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat 2761 gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga 2821 tagagaacgt atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc 2881 cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag 2941 cagagctcgt ttagtgaacc gtcagatcgc cactagtcga ctggatccgg taccatggag 3001 cagaagctga tctccgagga ggacctggga tccatggctg ccatccggaa gaaactggtg 3061 attgttggtg atggagcctg tggaaagaca tgcttgctca tagtcttcag caaggaccag 3121 ttcccagagg tgtatgtgcc cacagtgttt gagaactatg tggcagatat cgaggtggat 3181 ggaaagcagg tagagttggc tttgtgggac acagctgggc tggaagatta tgaccgcctg 3241 aggcccctct cctacccaga taccgatgtt atactgatgt gtttttccat cgacagccct 3301 gatagtttag aaaacatccc agaaaagtgg accccagaag tcaagcattt ctgtcccaac 3361 gtgcccatca tcctggttgg gaataagaag gatcttcgga atgatgagca cacaaggcgg 3421 gagctagcca agatgaagca ggagccggtg aaacctgaag aaggcagaga tatggcaaac 3481 aggattggcg cttttgggta catggagtgt tcagcaaaga ccaaagatgg agtgagagag 3541 gtttttgaaa tggctacgag agctgctctg caagctagac gtgggaagaa aaaatctggg 3601 tgccttgtct tgtgaaacct tgctggaagc acagccctta tgcggtgaat tctgcagata 3661 tccatcacac tggcggccgc tcgagatatc tagacccagc tttcttgtac aaagtggttg 3721 ataacatctc gagccataaa gatggttaat taacccaccc aagatctggc ctccgcgccg 3781 ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca cgtcagacga 3841 agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc 3901 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg 3961 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct 4021 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga 4081 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt 4141 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc 4201 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt 4261 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg 4321 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca 4381 aggtgggggg catggtgggc ggcaagaacc caaggtcttg aggccttcgc taatgcggga 4441 aagctcttat tcgggtgaga tgggctgggg caccatctgg ggaccctgac gtgaagtttg 4501 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg 4561 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg 4621 ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc 4681 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg 4741 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct 4801 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt 4861 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc 4921 agtgttagac tagtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagacg 4981 aagcttggta ccgagctcgg atctccaccc cgtaccggtc ctgcaggatt taaatggcgc 5041 gccgctagcg ctaccggact cagatctcga gctcaagctt cgaattctgc agtcgaattc 5101 accatgtcta gactggacaa gagcaaagtc ataaacggag ctctggaatt actcaatggt 5161 gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct 5221 accctgtact ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg 5281 ctggacaggc atcataccca cttctgcccc ctggaaggcg agtcatggca agactttctg 5341 cggaacaacg ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg 5401 catctcggca cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc 5461 ctgtgtcagc aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac 5521 tttacactgg gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag 5581 acacctacca ccgattctat gcccccactt ctgagacaag caattgagct gttcgaccgg 5641 cagggagccg aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa 5701 cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg acgattttga cttagacatg 5761 ctcccagccg atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac 5821 gattttgacc ttgacatgct ccccgggtaa ctaagtaagg atccgcggcc gcactagagg 5881 aattccgccc ctctcccccc cccccctctc cctccccccc ccctaacgtt actggccgaa 5941 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt 6001 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 6061 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 6121 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 6181 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 6241 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 6301 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 6361 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac 6421 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt 6481 accggtacgc gtgatgatcc accatggtga gcaagggcga ggagctgttc accggggtgg 6541 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 6601 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca 6661 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg 6721 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 6781 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 6841 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 6901 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 6961 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg 7021 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 7081 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca 7141 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 7201 gcatggacga gctgtacaag taagaattgt tgttgtttag tccctcccaa ttcgatatca 7261 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 7321 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 7381 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 7441 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 7501 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 7561 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 7621 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 7681 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 7741 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 7801 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 7861 atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat 7921 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt 7981 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt 8041 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa 8101 gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac 8161 acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca 8221 gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct 8281 gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc 8341 cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg 8401 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 8461 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 8521 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta 8581 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 8641 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 8701 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 8761 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 8821 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 8881 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 8941 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 9001 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 9061 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 9121 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 9181 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 9241 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 9301 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 9361 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 9421 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 9481 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 9541 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 9601 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 9661 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg 9721 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc 9781 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt 9841 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt 9901 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc 9961 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg 10021 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc 10081 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac 10141 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 10201 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 10261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 10321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 10381 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 10441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 10501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 10561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 10621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 10681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 10741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 10801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 10861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 10921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 10981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 11041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 11101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 11161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 11221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 11281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 11341 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 11401 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 11461 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 11521 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 11581 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 11641 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 11701 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 11761 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 11821 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 11881 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 11941 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 12001 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 12061 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 12121 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 12181 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 12241 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 12301 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 12361 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 12421 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 12481 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 12541 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgg //