pLenti6.2/V5-DEST

pLenti6.2/V5-DEST

pLenti6.2/V5-DEST载体基本信息：

pLenti6.2/V5-DEST载体质粒图谱和多克隆位点信息：

pLenti6.2/V5-DEST载体简介：

pLenti6.2/V5-DEST载体描述

The pLenti6.2/V5-DEST Gateway Vector is a Gateway-adapted ViraPower II lentiviral expression vector for lentiviral-based expression of a target gene in dividing and non-dividing mammalian cells. The vector has the CMV promoter for driving constitutive expression of the target gene and the PGK promoter for driving long-term, persistent expression of the Blasticidin stable selection marker.

pLenti6.2/V5-DEST载体优点

 Stable expression

 Long-term experiments

 High-throughput screening

 Accurate titer of functional virus (using Blasticidin method)

 Flexible and versatile Gateway recombination cloning technology

pLenti6.2/V5-DEST载体特征

 Human cytomegalovirus (CMV) immediate early promoter to control high-level expression of the gene of interest

 PKG Promoter for expression of Blasticidin selection marker

Gateway技术

The Gateway Technology is a universal cloning method that takes advantage of the site-specific recombination properties of bacteriophage lambda (Landy, 1989) to provide a rapid and highly efficient way to move your gene of interest into multiple vector systems. To express your gene of interest using

Gateway Technology, simply:

1. Generate entry clones containing your promoter and gene(s) of interest.

2. Generate an expression clone by performing an LR recombination reaction between the entry clone(s) and pcDNA6.2/V5-pL-DEST).

3. Transfect your expression clone into cells of your choice to transiently or stably express your gene of interest.

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate ORF Clone. The following table lists a variety of available destination vectors.

Additional materials required, available separately: Gateway entry clone, appropriate Gateway LR Clonase enzyme mix, and reaction buffer.

所需材料

在着手开始实验前你需要准备一下材料：

Gateway entry clone, appropriate Gateway LR Clonase enzyme mix, and reaction buffer.

• Purified plasmid DNA of your entry clone(s) (10 fmoles each)

• pcDNA6.2/V5-pL-DEST (20 fmoles)

• LR Clonase II Plus enzyme mix (keep at –20°C until immediately before use)

• 1X TE Buffer, pH 8.0 (10 mM Tris-HCl, pH 8.0, 1 mM EDTA)

• 2 μg/μL Proteinase K solution (supplied with the enzyme mix; thaw and keep on ice until use)

• Appropriate competent E. coli host and growth media for expression

• S.O.C. Medium

• LB agar plates containing 100 μg/mL ampicillin

进行LR重组反应

值得注意的事项：

If you use E. coli cells with a transformation efficiency of ≥1 × 108 cfu/μg, a typical LR reaction should give >5,000 colonies if the entire reaction is transformed and plated.

For multiple fragment reactions, typical numbers of colonies (per 10 μL LR reaction) are:

• 2-fragment recombination reaction: 2,000–15,000

• 3-fragment recombination reaction: 1,000–5,000

• 4-fragment recombination reaction: 50–500

Confirming the Expression Clone

The ccdB gene mutates at a very low frequency, resulting in a very low number of false positives. True expression clones will be ampicillin-resistant and chloramphenicol-sensitive. Transformants containing a plasmid with a mutated ccdB gene will be both ampicillin- and chloramphenicol-resistant.

To check your putative expression clone, test for growth on LB plates containing 30 μg/mL chloramphenicol. A true expression clone will not grow in the presence of chloramphenicol.

Sequencing To confirm that your gene of interest is in frame with the C-terminal V5 epitope, you may sequence your expression construct, if desired. We suggest using the following primer sequences. Refer to the diagram on page 6 for the location of the primer binding sites.

pLenti6.2/V5-DEST载体序列：

ORIGIN

    1 AATGTAGTCT TATGCAATAC TCTTGTAGTC TTGCAACATG GTAACGATGA GTTAGCAACA

   61 TGCCTTACAA GGAGAGAAAA AGCACCGTGC ATGCCGATTG GTGGAAGTAA GGTGGTACGA

  121 TCGTGCCTTA TTAGGAAGGC AACAGACGGG TCTGACATGG ATTGGACGAA CCACTGAATT

  181 GCCGCATTGC AGAGATATTG TATTTAAGTG CCTAGCTCGA TACATAAACG GGTCTCTCTG

  241 GTTAGACCAG ATCTGAGCCT GGGAGCTCTC TGGCTAACTA GGGAACCCAC TGCTTAAGCC

  301 TCAATAAAGC TTGCCTTGAG TGCTTCAAGT AGTGTGTGCC CGTCTGTTGT GTGACTCTGG

  361 TAACTAGAGA TCCCTCAGAC CCTTTTAGTC AGTGTGGAAA ATCTCTAGCA GTGGCGCCCG

  421 AACAGGGACT TGAAAGCGAA AGGGAAACCA GAGGAGCTCT CTCGACGCAG GACTCGGCTT

  481 GCTGAAGCGC GCACGGCAAG AGGCGAGGGG CGGCGACTGG TGAGTACGCC AAAAATTTTG

  541 ACTAGCGGAG GCTAGAAGGA GAGAGATGGG TGCGAGAGCG TCAGTATTAA GCGGGGGAGA

  601 ATTAGATCGC GATGGGAAAA AATTCGGTTA AGGCCAGGGG GAAAGAAAAA ATATAAATTA

  661 AAACATATAG TATGGGCAAG CAGGGAGCTA GAACGATTCG CAGTTAATCC TGGCCTGTTA

  721 GAAACATCAG AAGGCTGTAG ACAAATACTG GGACAGCTAC AACCATCCCT TCAGACAGGA

  781 TCAGAAGAAC TTAGATCATT ATATAATACA GTAGCAACCC TCTATTGTGT GCATCAAAGG

  841 ATAGAGATAA AAGACACCAA GGAAGCTTTA GACAAGATAG AGGAAGAGCA AAACAAAAGT

  901 AAGACCACCG CACAGCAAGC GGCCGCTGAT CTTCAGACCT GGAGGAGGAG ATATGAGGGA

  961 CAATTGGAGA AGTGAATTAT ATAAATATAA AGTAGTAAAA ATTGAACCAT TAGGAGTAGC

 1021 ACCCACCAAG GCAAAGAGAA GAGTGGTGCA GAGAGAAAAA AGAGCAGTGG GAATAGGAGC

 1081 TTTGTTCCTT GGGTTCTTGG GAGCAGCAGG AAGCACTATG GGCGCAGCGT CAATGACGCT

 1141 GACGGTACAG GCCAGACAAT TATTGTCTGG TATAGTGCAG CAGCAGAACA ATTTGCTGAG

 1201 GGCTATTGAG GCGCAACAGC ATCTGTTGCA ACTCACAGTC TGGGGCATCA AGCAGCTCCA

 1261 GGCAAGAATC CTGGCTGTGG AAAGATACCT AAAGGATCAA CAGCTCCTGG GGATTTGGGG

 1321 TTGCTCTGGA AAACTCATTT GCACCACTGC TGTGCCTTGG AATGCTAGTT GGAGTAATAA

 1381 ATCTCTGGAA CAGATTTGGA ATCACACGAC CTGGATGGAG TGGGACAGAG AAATTAACAA

 1441 TTACACAAGC TTAATACACT CCTTAATTGA AGAATCGCAA AACCAGCAAG AAAAGAATGA

 1501 ACAAGAATTA TTGGAATTAG ATAAATGGGC AAGTTTGTGG AATTGGTTTA ACATAACAAA

 1561 TTGGCTGTGG TATATAAAAT TATTCATAAT GATAGTAGGA GGCTTGGTAG GTTTAAGAAT

 1621 AGTTTTTGCT GTACTTTCTA TAGTGAATAG AGTTAGGCAG GGATATTCAC CATTATCGTT

 1681 TCAGACCCAC CTCCCAACCC CGAGGGGACC CGACAGGCCC GAAGGAATAG AAGAAGAAGG

 1741 TGGAGAGAGA GACAGAGACA GATCCATTCG ATTAGTGAAC GGATCTCGAC GGTATCGATA

 1801 AGCTTGGGAG TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA

 1861 CGACCCCCGC CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC CAATAGGGAC

 1921 TTTCCATTGA CGTCAATGGG TGGAGTATTT ACGGTAAACT GCCCACTTGG CAGTACATCA

 1981 AGTGTATCAT ATGCCAAGTA CGCCCCCTAT TGACGTCAAT GACGGTAAAT GGCCCGCCTG

 2041 GCATTATGCC CAGTACATGA CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT

 2101 AGTCATCGCT ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG

 2161 GTTTGACTCA CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA GTTTGTTTTG

 2221 GCACCAAAAT CAACGGGACT TTCCAAAATG TCGTAACAAC TCCGCCCCAT TGACGCAAAT

 2281 GGGCGGTAGG CGTGTACGGT GGGAGGTCTA TATAAGCAGA GCTCGTTTAG TGAACCGTCA

 2341 GATCGCCTGG AGACGCCATC CACGCTGTTT TGACCTCCAT AGAAGACACC GACTCTAGAG

 2401 GATCCACTAG TCCAGTGTGG TGGAATTCTG CAGATATCAA CAAGTTTGTA CAAAAAAGCT

 2461 GAACGAGAAA CGTAAAATGA TATAAATATC AATATATTAA ATTAGATTTT GCATAAAAAA

 2521 CAGACTACAT AATACTGTAA AACACAACAT ATCCAGTCAC TATGGCGGCC GCATTAGGCA

 2581 CCCCAGGCTT TACACTTTAT GCTTCCGGCT CGTATAATGT GTGGATTTTG AGTTAGGATC

 2641 CGGCGAGATT TTCAGGAGCT AAGGAAGCTA AAATGGAGAA AAAAATCACT GGATATACCA

 2701 CCGTTGATAT ATCCCAATGG CATCGTAAAG AACATTTTGA GGCATTTCAG TCAGTTGCTC

 2761 AATGTACCTA TAACCAGACC GTTCAGCTGG ATATTACGGC CTTTTTAAAG ACCGTAAAGA

 2821 AAAATAAGCA CAAGTTTTAT CCGGCCTTTA TTCACATTCT TGCCCGCCTG ATGAATGCTC

 2881 ATCCGGAATT CCGTATGGCA ATGAAAGACG GTGAGCTGGT GATATGGGAT AGTGTTCACC

 2941 CTTGTTACAC CGTTTTCCAT GAGCAAACTG AAACGTTTTC ATCGCTCTGG AGTGAATACC

 3001 ACGACGATTT CCGGCAGTTT CTACACATAT ATTCGCAAGA TGTGGCGTGT TACGGTGAAA

 3061 ACCTGGCCTA TTTCCCTAAA GGGTTTATTG AGAATATGTT TTTCGTCTCA GCCAATCCCT

 3121 GGGTGAGTTT CACCAGTTTT GATTTAAACG TGGCCAATAT GGACAACTTC TTCGCCCCCG

 3181 TTTTCACCAT GGGCAAATAT TATACGCAAG GCGACAAGGT GCTGATGCCG CTGGCGATTC

 3241 AGGTTCATCA TGCCGTCTGT GATGGCTTCC ATGTCGGCAG AATGCTTAAT GAATTACAAC

 3301 AGTACTGCGA TGAGTGGCAG GGCGGGGCGT AAAGATCTGG ATCCGGCTTA CTAAAAGCCA

 3361 GATAACAGTA TGCGTATTTG CGCGCTGATT TTTGCGGTAT AAGAATATAT ACTGATATGT

 3421 ATACCCGAAG TATGTCAAAA AGAGGTGTGC TATGAAGCAG CGTATTACAG TGACAGTTGA

 3481 CAGCGACAGC TATCAGTTGC TCAAGGCATA TATGATGTCA ATATCTCCGG TCTGGTAAGC

 3541 ACAACCATGC AGAATGAAGC CCGTCGTCTG CGTGCCGAAC GCTGGAAAGC GGAAAATCAG

 3601 GAAGGGATGG CTGAGGTCGC CCGGTTTATT GAAATGAACG GCTCTTTTGC TGACGAGAAC

 3661 AGGGACTGGT GAAATGCAGT TTAAGGTTTA CACCTATAAA AGAGAGAGCC GTTATCGTCT

 3721 GTTTGTGGAT GTACAGAGTG ATATTATTGA CACGCCCGGG CGACGGATGG TGATCCCCCT

 3781 GGCCAGTGCA CGTCTGCTGT CAGATAAAGT CTCCCGTGAA CTTTACCCGG TGGTGCATAT

 3841 CGGGGATGAA AGCTGGCGCA TGATGACCAC CGATATGGCC AGTGTGCCGG TCTCCGTTAT

 3901 CGGGGAAGAA GTGGCTGATC TCAGCCACCG CGAAAATGAC ATCAAAAACG CCATTAACCT

 3961 GATGTTCTGG GGAATATAAA TGTCAGGCTC CGTTATACAC AGCCAGTCTG CAGGTCGACC

 4021 ATAGTGACTG GATATGTTGT GTTTTACAGT ATTATGTAGT CTGTTTTTTA TGCAAAATCT

 4081 AATTTAATAT ATTGATATTT ATATCATTTT ACGTTTCTCG TTCAGCTTTC TTGTACAAAG

 4141 TGGTTGATAT CCAGCACAGT GGCGGCCGCT CGAGTCTAGA GGGCCCGCGG TTCGAAGGTA

 4201 AGCCTATCCC TAACCCTCTC CTCGGTCTCG ATTCTACGCG TACCGGTTAG TAATGACGAT

 4261 GGTACCTACC GGGTAGGGGA GGCGCTTTTC CCAAGGCAGT CTGGAGCATG CGCTTTAGCA

 4321 GCCCCGCTGG GCACTTGGCG CTACACAAGT GGCCTCTGGC CTCGCACACA TTCCACATCC

 4381 ACCGGTAGGC GCCAACCGGC TCCGTTCTTT GGTGGCCCCT TCGCGCCACC TTCTACTCCT

 4441 CCCCTAGTCA GGAAGTTCCC CCCCGCCCCG CAGCTCGCGT CGTGCAGGAC GTGACAAATG

 4501 GAAGTAGCAC GTCTCACTAG TCTCGTGCAG ATGGACAGCA CCGCTGAGCA ATGGAAGCDG

 4561 GTAGGCCTTT GGGGCAGCGG CCAATAGCAG CTTTGCTCCT TCGCTTTCTG GGCTCAGAGG

 4621 CTGGGAAGGG GTGGGTCCGG GGGCGGGCTC AGGGGCGGGC TCAGGGGCGG GGCGGGCGCC

 4681 CGAAGTCCTC CGGAGGCCCG GCATTCTGCA CGCTTCAAAA GCGCACGTCT GCCGCGCTGT

 4741 TCTCCTCTTC CTCATCTCCG GGCCTTTCGA CTCTAGACAC GTGTTGACAA TTAATCATCG

 4801 GCATAGTATA TCGGCATAGT ATAATACGAC AAGGTGAGGA ACTAAACCAT GGCCAAGCCT

 4861 TTGTCTCAAG AAGAATCCAC CCTCATTGAA AGAGCAACGG CTACAATCAA CAGCATCCCC

 4921 ATCTCTGAAG ACTACAGCGT CGCCAGCGCA GCTCTCTCTA GCGACGGCCG CATCTTCACT

 4981 GGTGTCAATG TATATCATTT TACTGGGGGA CCTTGTGCAG AACTCGTGGT GCTGGGCACT

 5041 GCTGCTGCTG CGGCAGCTGG CAACCTGACT TGTATCGTCG CGATCGGAAA TGAGAACAGG

 5101 GGCATCTTGA GCCCCTGCGG ACGGTGCCGA CAGGTGCTTC TCGATCTGCA TCCTGGGATC

 5161 AAAGCCATAG TGAAGGACAG TGATGGACAG CCGACGGCAG TTGGGATTCG TGAATTGCTG

 5221 CCCTCTGGTT ATGTGTGGGA GGGCTAAGCA CAATTCGAGC TCGGTACCTT TAAGACCAAT

 5281 GACTTACAAG GCAGCTGTAG ATCTTAGCCA CTTTTTAAAA GAAAAGGGGG GACTGGAAGG

 5341 GCTAATTCAC TCCCAACGAA GACAAGATCT GCTTTTTGCT TGTACTGGGT CTCTCTGGTT

 5401 AGACCAGATC TGAGCCTGGG AGCTCTCTGG CTAACTAGGG AACCCACTGC TTAAGCCTCA

 5461 ATAAAGCTTG CCTTGAGTGC TTCAAGTAGT GTGTGCCCGT CTGTTGTGTG ACTCTGGTAA

 5521 CTAGAGATCC CTCAGACCCT TTTAGTCAGT GTGGAAAATC TCTAGCAGTA GTAGTTCATG

 5581 TCATCTTATT ATTCAGTATT TATAACTTGC AAAGAAATGA ATATCAGAGA GTGAGAGGAA

 5641 CTTGTTTATT GCAGCTTATA ATGGTTACAA ATAAAGCAAT AGCATCACAA ATTTCACAAA

 5701 TAAAGCATTT TTTTCACTGC ATTCTAGTTG TGGTTTGTCC AAACTCATCA ATGTATCTTA

 5761 TCATGTCTGG CTCTAGCTAT CCCGCCCCTA ACTCCGCCCA TCCCGCCCCT AACTCCGCCC

 5821 AGTTCCGCCC ATTCTCCGCC CCATGGCTGA CTAATTTTTT TTATTTATGC AGAGGCCGAG

 5881 GCCGCCTCGG CCTCTGAGCT ATTCCAGAAG TAGTGAGGAG GCTTTTTTGG AGGCCTAGGG

 5941 ACGTACCCAA TTCGCCCTAT AGTGAGTCGT ATTACGCGCG CTCACTGGCC GTCGTTTTAC

 6001 AACGTCGTGA CTGGGAAAAC CCTGGCGTTA CCCAACTTAA TCGCCTTGCA GCACATCCCC

 6061 CTTTCGCCAG CTGGCGTAAT AGCGAAGAGG CCCGCACCGA TCGCCCTTCC CAACAGTTGC

 6121 GCAGCCTGAA TGGCGAATGG GACGCGCCCT GTAGCGGCGC ATTAAGCGCG GCGGGTGTGG

 6181 TGGTTACGCG CAGCGTGACC GCTACACTTG CCAGCGCCCT AGCGCCCGCT CCTTTCGCTT

 6241 TCTTCCCTTC CTTTCTCGCC ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA AATCGGGGGC

 6301 TCCCTTTAGG GTTCCGATTT AGTGCTTTAC GGCACCTCGA CCCCAAAAAA CTTGATTAGG

 6361 GTGATGGTTC ACGTAGTGGG CCATCGCCCT GATAGACGGT TTTTCGCCCT TTGACGTTGG

 6421 AGTCCACGTT CTTTAATAGT GGACTCTTGT TCCAAACTGG AACAACACTC AACCCTATCT

 6481 CGGTCTATTC TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG

 6541 AGCTGATTTA ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTAGG

 6601 TGGCACTTTT CGGGGAAATG TGCGCGGAAC CCCTATTTGT TTATTTTTCT AAATACATTC

 6661 AAATATGTAT CCGCTCATGA GACAATAACC CTGATAAATG CTTCAATAAT ATTGAAAAAG

 6721 GAAGAGTATG AGTATTCAAC ATTTCCGTGT CGCCCTTATT CCCTTTTTTG CGGCATTTTG

 6781 CCTTCCTGTT TTTGCTCACC CAGAAACGCT GGTGAAAGTA AAAGATGCTG AAGATCAGTT

 6841 GGGTGCACGA GTGGGTTACA TCGAACTGGA TCTCAACAGC GGTAAGATCC TTGAGAGTTT

 6901 TCGCCCCGAA GAACGTTTTC CAATGATGAG CACTTTTAAA GTTCTGCTAT GTGGCGCGGT

 6961 ATTATCCCGT ATTGACGCCG GGCAAGAGCA ACTCGGTCGC CGCATACACT ATTCTCAGAA

 7021 TGACTTGGTT GAGTACTCAC CAGTCACAGA AAAGCATCTT ACGGATGGCA TGACAGTAAG

 7081 AGAATTATGC AGTGCTGCCA TAACCATGAG TGATAACACT GCGGCCAACT TACTTCTGAC

 7141 AACGATCGGA GGACCGAAGG AGCTAACCGC TTTTTTGCAC AACATGGGGG ATCATGTAAC

 7201 TCGCCTTGAT CGTTGGGAAC CGGAGCTGAA TGAAGCCATA CCAAACGACG AGCGTGACAC

 7261 CACGATGCCT GTAGCAATGG CAACAACGTT GCGCAAACTA TTAACTGGCG AACTACTTAC

 7321 TCTAGCTTCC CGGCAACAAT TAATAGACTG GATGGAGGCG GATAAAGTTG CAGGACCACT

 7381 TCTGCGCTCG GCCCTTCCGG CTGGCTGGTT TATTGCTGAT AAATCTGGAG CCGGTGAGCG

 7441 TGGGTCTCGC GGTATCATTG CAGCACTGGG GCCAGATGGT AAGCCCTCCC GTATCGTAGT

 7501 TATCTACACG ACGGGGAGTC AGGCAACTAT GGATGAACGA AATAGACAGA TCGCTGAGAT

 7561 AGGTGCCTCA CTGATTAAGC ATTGGTAACT GTCAGACCAA GTTTACTCAT ATATACTTTA

 7621 GATTGATTTA AAACTTCATT TTTAATTTAA AAGGATCTAG GTGAAGATCC TTTTTGATAA

 7681 TCTCATGACC AAAATCCCTT AACGTGAGTT TTCGTTCCAC TGAGCGTCAG ACCCCGTAGA

 7741 AAAGATCAAA GGATCTTCTT GAGATCCTTT TTTTCTGCGC GTAATCTGCT GCTTGCAAAC

 7801 AAAAAAACCA CCGCTACCAG CGGTGGTTTG TTTGCCGGAT CAAGAGCTAC CAACTCTTTT

 7861 TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA GATACCAAAT ACTGTTCTTC TAGTGTAGCC

 7921 GTAGTTAGGC CACCACTTCA AGAACTCTGT AGCACCGCCT ACATACCTCG CTCTGCTAAT

 7981 CCTGTTACCA GTGGCTGCTG CCAGTGGCGA TAAGTCGTGT CTTACCGGGT TGGACTCAAG

 8041 ACGATAGTTA CCGGATAAGG CGCAGCGGTC GGGCTGAACG GGGGGTTCGT GCACACAGCC

 8101 CAGCTTGGAG CGAACGACCT ACACCGAACT GAGATACCTA CAGCGTGAGC TATGAGAAAG

 8161 CGCCACGCTT CCCGAAGGGA GAAAGGCGGA CAGGTATCCG GTAAGCGGCA GGGTCGGAAC

 8221 AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG AAACGCCTGG TATCTTTATA GTCCTGTCGG

 8281 GTTTCGCCAC CTCTGACTTG AGCGTCGATT TTTGTGATGC TCGTCAGGGG GGCGGAGCCT

 8341 ATGGAAAAAC GCCAGCAACG CGGCCTTTTT ACGGTTCCTG GCCTTTTGCT GGCCTTTTGC

 8401 TCACATGTTC TTTCCTGCGT TATCCCCTGA TTCTGTGGAT AACCGTATTA CCGCCTTTGA

 8461 GTGAGCTGAT ACCGCTCGCC GCAGCCGAAC GACCGAGCGC AGCGAGTCAG TGAGCGAGGA

 8521 AGCGGAAGAG CGCCCAATAC GCAAACCGCC TCTCCCCGCG CGTTGGCCGA TTCATTAATG

 8581 CAGCTGGCAC GACAGGTTTC CCGACTGGAA AGCGGGCAGT GAGCGCAACG CAATTAATGT

 8641 GAGTTAGCTC ACTCATTAGG CACCCCAGGC TTTACACTTT ATGCTTCCGG CTCGTATGTT

 8701 GTGTGGAATT GTGAGCGGAT AACAATTTCA CACAGGAAAC AGCTATGACC ATGATTACGC

 8761 CAAGCGCGCA ATTAACCCTC ACTAAAGGGA ACAAAAGCTG GAGCTGCAAG CTT

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