质粒名称: | pSBtet-GP |
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出品公司: | 爱迪基因 |
目录编号: | 60495 |
存储实验室: | Eric Kowarz |
载体骨架基本信息 |
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载体名称: | pUC19 |
载体抗性: | 氨苄青霉素 |
空载体大小: | 2021 bp |
载体类型: | 哺乳动物表达;转座子 |
筛选标记: | 嘌呤霉素 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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启动子 | TCE & RPBSA |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 测序引物 | CCTGGAGCCAATTCCAACTCT |
3' 测序引物 | CACTGCATTCTTGTTGTGGTT |
文献引用方法 |
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材料和方法部分: | pSBtet-GP was a gift from Eric Kowarz (Addgene plasmid # 60495 ; http://n2t.net/addgene:60495 ; RRID:Addgene_60495) |
参考文献部分: | Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Kowarz E, Loescher D, Marschalek R. Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821. 10.1002/biot.201400821 PubMed 25650551 |
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pSBtetGP载体基本信息
pSBtetGP载体图谱质粒图谱和多克隆位点信息
pSBtetGP载体简介
pSBtetGP载体序列
LOCUS pSBtet-GP 8021 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION SB-transposon with inducible SfiI cloning site for GOI (contains firefly luciferase) and constitutive expression of GFP, rtTA and puromycin resistance gene. ACCESSION . VERSION . KEYWORDS pSBtet-GP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8021) AUTHORS Kowarz E, Loescher D, Marschalek R TITLE Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. JOURNAL Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821. PUBMED 25650551 REFERENCE 2 (bases 1 to 8021) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8021 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 295..607 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 303..321 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 339..357 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 374..392 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 410..428 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 446..464 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 481..499 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 517..535 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 580..604 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 680..689 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 686..2338 /codon_start=1 /product="firefly luciferase" /label=luciferase /note="synthetic luc2 version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" polyA_signal 2374..2495 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2385..2404) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2439..2458 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS 3218..3934 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3218..3934 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3263..3284) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3524..3543) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3871..3892 /label=EGFP-C /note="EGFP, forward primer" CDS 3944..4000 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 4001..4744 /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 KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" CDS 4751..4807 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 4808..5407 /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(4808..4827) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5304..5324 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(5462..5479) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5468..5692 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6018..6035) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6189..6777) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6269..6288) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6948..7808) /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 7571..7590 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7809..7913) /gene="bla" /label=AmpR promoter ORIGIN 1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact 61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta 121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta 181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt 241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agacgagttt 301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg 361 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt 421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta 481 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta 541 tgtcgattag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca 601 gatcgcctgg agccaattcc aactcttttg ccttatacct actttccgta ccacttccta 661 ccctcgaaag gcctctgagg ccaccatgga agatgccaaa aacattaaga agggcccagc 721 gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg 781 ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac 841 ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct 901 gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt 961 gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg 1021 cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg 1081 gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat 1141 ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt 1201 gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat 1261 cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca 1321 ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat 1381 ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac 1441 gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct 1501 attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt 1561 tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat 1621 cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca 1681 cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac 1741 ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa 1801 ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt 1861 ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat 1921 cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt 1981 cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc 2041 cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct 2101 gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac 2161 catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct 2221 gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc 2281 ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaagg 2341 cctgtcaggc caagcttcca tcgatagaca tgataagata cattgatgag tttggacaaa 2401 ccacaacaag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt 2461 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta 2521 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat 2581 gtggtaccta ggcgagaccc tgtctcacaa aataaagtaa gcccggactg agtgcggaaa 2641 ggcgggcctg gcgggtctgg tctccccatg cgggccacca gaggccctgc agccttcagt 2701 cgcttgaagg ggtaatggcg cttccactca caaacatggc ggacagagcg tgtgaacgag 2761 atgaacagcc cctcaaaaat atggccgccg aggctggacg gccgtgcccc agcagcaccg 2821 cctccgcgcc ccacgtgatc tctcgccggg cacagcgctg accgcggagg tccaaccgga 2881 agaatgtccg gattggacat tcggaagagg gcccgccttc cctggggaat ctctgcgcac 2941 gcgcagaacg cttcgaccaa tgaaaacaca ggaagccgtc cgcgcaaccg cgttgcgtca 3001 cttctgccgc ccctgtttca aggtatatag ccgtagacgg aacttcgcct ttctctcggc 3061 cttagcgcca tttttttggg tgagtgtttt ttggttcctg cgttgggatt ccgtgtacaa 3121 tccatagaca tctgacctcg gcacttagca tcatcacagc aaactaactg tagcctttct 3181 ctctttccct gtagaaacct ctgcacctga ggccaccatg ttgagcaagg gcgaggagct 3241 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt 3301 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat 3361 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg 3421 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc 3481 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa 3541 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg 3601 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag 3661 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat 3721 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc 3781 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct 3841 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 3901 cgggatcact ctcggcatgg acgagctgta caagggcagt ggagctacta acttcagcct 3961 gctgaagcag gctggtgacg tcgaggagaa tcctggcccc atgtctagac tggacaagag 4021 caaagtcata aacggcgctc tggaattact caatggagtc ggtatcgaag gcctgacgac 4081 aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa 4141 caagcgggcc ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt 4201 ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcattccg 4261 ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga 4321 gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct 4381 ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga 4441 ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc 4501 cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct 4561 tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg 4621 gccggccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga 4681 ctttgacctt gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc 4741 cgggtccggc gctactaact tcagcctgct gaagcaggct ggtgacgtcg aggagaatcc 4801 tggtcccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag 4861 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga 4921 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg 4981 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac 5041 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt 5101 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc 5161 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg 5221 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc 5281 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt 5341 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg 5401 tgcctgattc gaaggcctgt cgtgaagctt ggggatcaat tctctagagc tcgctgatca 5461 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 5521 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 5581 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 5641 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 5701 gcggaaagaa ccagctgggc ctactagcta ctcgggaccc cttaccgaaa catcgccgca 5761 ttctgcagag gagtcgagtg tatgtaaact tctgacccac tgggaatgtg atgaaagaaa 5821 taaaagctga aatgaatcat tctctctact attattctga tatttcacat tcttaaaata 5881 aagtggtgat cctaactgac ctaagacagg gaatttttac taggattaaa tgtcaggaat 5941 tgtgaaaaag tgagtttaaa tgtatttggc taaggtgtat gtaaacttcc gacttcaact 6001 gtatagggat ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 6061 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 6121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 6181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 6241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 6301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 6361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 6421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 6481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 6541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 6601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 6661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 6721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 6781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 6841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 7021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 7081 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 7141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 7201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 7261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 7321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 7381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 7441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 7501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 7561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 7621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 7681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 7741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 7801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 7861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 7921 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 7981 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat t //