LOCUS Exported 10490 bp ds-DNA circular SYN 23-SEP-2022 DEFINITION 3rd generation lentiviral plasmid for expression of EGFP Firefly Luciferase fusion protein in mammalian cells. ACCESSION . VERSION . KEYWORDS pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10490) AUTHORS Lorenzatti Hiles G, Cates AL, El-Sawy L, Day KC, Broses LJ, Han AL, Briggs HL, Emamdjomeh A, Chou A, Abel EV, Liebert M, Palmbos PL, Udager AM, Keller ET, Day ML TITLE A surgical orthotopic approach for studying the invasive progression of human bladder cancer. JOURNAL Nat Protoc. 2019 Mar;14(3):738-755. doi: 10.1038/s41596-018-0112-8. PUBMED 30683938 REFERENCE 2 (bases 1 to 10490) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10490 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(35..52) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(206..794) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(286..305) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(965..1825) /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 1588..1607 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1826..1930) /gene="bla" /label=AmpR promoter rep_origin complement(1956..2411) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2093..2114) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2305..2324 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2538..2560 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2552..2569 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2553..2569 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2579..2598 /label=T7 /note="T7 promoter, forward primer" promoter 2579..2597 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" rep_origin 2618..2753 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(2672..2691) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 2793..2914 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2804..2823) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2858..2877 /label=SV40pA-R /note="SV40 polyA, reverse primer" LTR 2986..3219 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" misc_feature 3299..3887 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 3405..3416 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 3814..3834 /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3955..4554) /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(4038..4058) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 4535..4554 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(4576..5086) /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind complement(4639..4657) /label=hPGK-F /note="Human PGK promoter, forward primer" CDS complement(5146..5187) /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" protein_bind 5240..5264 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS complement(5281..6930) /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind 6846..6864 /label=LucNrev /note="Firefly luciferase, reverse primer" CDS complement(6937..7653) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS complement(6937..7653) /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(6979..7000) /label=EGFP-C /note="EGFP, forward primer" primer_bind 7328..7347 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7587..7608 /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(7674..7698) /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind 7739..7757 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 7760..7778 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" promoter complement(7780..7983) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(7809..7829) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 8309..8426 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(8693..8737) /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 8922..9155 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 9648..9773 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" LTR 9820..10000 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(10001..10227) /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" primer_bind complement(10255..10275) /label=T3 /note="T3 promoter, forward primer" promoter complement(10255..10273) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(10294..10310) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10294..10310) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10307..10329) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10318..10334 /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(10342..10372) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10387..10408 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 1921 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt 1981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 2041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg 2101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 2161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg 2221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 2281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct 2341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct 2401 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 2461 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 2521 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta 2581 atacgactca ctatagggcg aattgggtac gtccctaggc ctccaaaaaa gcctcctcac 2641 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt 2701 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga 2761 gttaggggcg ggatagctag agccagacat gataagatac attgatgagt ttggacaaac 2821 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 2881 atttgtaacc attataagct gcaataaaca agttcctctc actctctgat attcatttct 2941 ttgcaagtta taaatactga ataataagat gacatgaact actactgcta gagattttcc 3001 acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac 3061 acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt 3121 agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta caagcaaaaa 3181 gcagatcttg tcttcgttgg gagtgaatta gcccttccag tccccccttt tcttttaaaa 3241 agtggctaag atctacagct gccttgtaag tcattggtct taaaggtacc atcgccaggc 3301 ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga 3361 ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg 3421 acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag cagccatgga 3481 aaggacgtca gcttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct 3541 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa 3601 gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg 3661 tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa 3721 agagacagca accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata 3781 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta 3841 agaataccag tcaatctttc acaaattttg taatccagag gttgattgtc gacgtaccga 3901 tgcatggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg ggcgtcaggc 3961 accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt 4021 gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa 4081 ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag 4141 acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt 4201 gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct 4261 caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg 4321 cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag 4381 cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg 4441 atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc 4501 cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatggtaag 4561 ctccggtgga tccccctggg gagagaggtc ggtgattcgg tcaacgaggg agccgactgc 4621 cgacgtgcgc tccggaggct tgcagaatgc ggaacaccgc gcgggcagga acagggccca 4681 cactaccgcc ccacaccccg cctcccgcac cgccccttcc cggccgctgc tctcggcgcg 4741 ccctgctgag cagccgctat tggccacagc ccatcgcggt cggcgcgctg ccattgctcc 4801 ctggcgctgt ccgtctgcga gggtactagt gagacgtgcg gcttccgttt gtcacgtccg 4861 gcacgccgcg aaccgcaagg aaccttcccg acttaggggc ggagcaggaa gcgtcgccgg 4921 ggggcccaca agggtagcgg cgaagatccg ggtgacgctg cgaacggacg tgaagaatgt 4981 gcgagaccca gggtcggcgc cgctgcgttt cccggaacca cgcccagagc agccgcgtcc 5041 ctgcgcaaac ccagggctgc cttggaaaag gcgcaacccc aaccccctcg actaactgac 5101 acacattcca cagaattaat tccaaactca ttactaaccg gtacgcgtag aatcgagacc 5161 gaggagaggg ttagggatag gcttaccttc gaaccgcggg ccctctagac tcgagcggcc 5221 gccactgtgc tggatatcaa ccactttgta caagaaagct gggtcggcgc gcccaccctt 5281 cacggcgatc tttccgccct tcttggcctt tatgaggatc tctctgattt ttcttgcgtc 5341 gagttttccg gtaagacctt tcggtacttc gtccacaaac acaactcctc cgcgcaactt 5401 tttcgcggtt gttacttgac tggcgacgta atccacgatc tctttttccg tcatcgtctt 5461 tccgtgctcc aaaacaacaa cggcggcggg aagttcaccg gcgtcatcgt cgggaagacc 5521 tgcgacacct gcgtcgaaga tgttggggtg ttggagcaag atggattcca attcagcggg 5581 agccacctga tagcctttgt acttaatcag agacttcagg cggtcaacga tgaagaagtg 5641 ttcgtcttcg tcccagtaag ctatgtctcc agaatgtagc catccatcct tgtcaatcaa 5701 ggcgttggtc gcttccggat tgtttacata accggacata atcataggac ctctcacaca 5761 cagttcgcct ctttgattaa cgcccagcgt tttcccggta tccagatcca caaccttcgc 5821 ttcaaaaaat ggaacaactt taccgaccgc gcccggttta tcatccccct cgggtgtaat 5881 cagaatagct gatgtagtct cagtgagccc atatccttgc ctgatacctg gcagatggaa 5941 cctcttggca accgcttccc cgacttcctt agagagggga gcgccaccag aagcaatttc 6001 gtgtaaatta gataaatcgt atttgtcaat cagagtgctt ttggcgaaga aggagaatag 6061 ggttggcacc agcagcgcac tttgaatctt gtaatcctga aggctcctca gaaacagctc 6121 ttcttcaaat ctatacatta agacgactcg aaatccacat atcaaatatc cgagtgtagt 6181 aaacattcca aaaccgtgat ggaatggaac aacacttaaa atcgcagtat ccggaatgat 6241 ttgattgcca aaaataggat ctctggcatg cgagaatctc acgcaggcag ttctatgagg 6301 cagagcgaca cctttaggca gaccagtaga tccagaggag ttcatgatca gtgcaattgt 6361 cttgtcccta tcgaaggact ctggcacaaa atcgtattca ttaaaaccgg gaggtagatg 6421 agatgtgacg aacgtgtaca tcgactgaaa tccctggtaa tccgttttag aatccatgat 6481 aataattttt tggatgattg ggagcttttt ttgcacgttc aaaatttttt gcaacccctt 6541 tttggaaacg aacaccacgg taggctgcga aatgcccata ctgttgagca attcacgttc 6601 attataaatg tcgttcgcgg gcgcaactgc aactccgata aataacgcgc ccaacaccgg 6661 cataaagaat tgaagagagt tttcactgca tacgacgatt ctgtgatttg tattcagccc 6721 atatcgtttc atagcttctg ccaaccgaac ggacatttcg aagtactcag cgtaagtgat 6781 gtccacctcg atatgtgcat ctgtaaaagc aattgttcca ggaaccaggg cgtatctctt 6841 catagcctta tgcagttgct ctccagcggt tccatcttcc agcggataga atggcgccgg 6901 gcctttcttt atgtttttgg cgtcttccat cccggacttg tacagctcgt ccatgccgag 6961 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg 7021 gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc 7081 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat 7141 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata 7201 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt 7261 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc 7321 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc 7381 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa 7441 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc 7501 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc 7561 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac 7621 cccggtgaac agctcctcgc ccttgctcac catggtgaag ggggcggccg cggagcctgc 7681 ttttttgtac aaacttgttg atatctgcag aattccacca cactggacta gtcgacgatc 7741 tctatcactg atagggagat ctctatcact gatagggaga gctctgctta tatagacctc 7801 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg 7861 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact 7921 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat 7981 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa 8041 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg 8101 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc 8161 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat 8221 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta 8281 tgtaagcgga actcccaagc ttatcgataa aattttgaat ttttgtaatt tgtttttgta 8341 attctttagt ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc 8401 ccccaatccc cccttttctt ttaaaagtta accgataccg tcgagatccg ttcactaatc 8461 gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg 8521 gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc 8581 tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca 8641 ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg 8701 cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt 8761 caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg 8821 tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag 8881 cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta 8941 ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt 9001 gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac 9061 cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc 9121 ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct 9181 gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt 9241 tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga 9301 tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct 9361 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact 9421 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc 9481 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct 9541 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt 9601 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac 9661 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc 9721 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct 9781 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg 9841 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta 9901 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag 9961 agagctccca ggctcagatc tggtctaacc agagagaccc gtttatgtat cgagctaggc 10021 acttaaatac aatatctctg caatgcggca attcagtggt tcgtccaatc catgtcagac 10081 ccgtctgttg ccttcctaat aaggcacgat cgtaccacct tacttccacc aatcggcatg 10141 cacggtgctt tttctctcct tgtaaggcat gttgctaact catcgttacc atgttgcaag 10201 actacaagag tattgcataa gactacatta agcttgcagc tccagctttt gttcccttta 10261 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 10321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 10381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 10441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga //