| 出品公司: | Addgene |
|---|---|
| 试剂名称: | pUMVC |
| 质粒类型: | 哺乳动物载体；逆病毒包装载体；双质粒包装系统 |
| 高拷贝/低拷贝: | 低拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | CMV |
| 载体大小: | 9287 bp |
| 5' 测序引物及序列: | CMV fwd 5’CGCAAATGGGCGGTAGGCGTG 3’ |
| 3' 测序引物及序列: | -- |
| 载体标签: | 无 |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | 无 |
| 克隆菌株: | DH5α等 |
| 宿主细胞（系）: | 包装细胞系如293T |
| 备注: | 逆病毒包装载体pUMVC需要与信封质粒 pCMV-VSV-G 或者 pCMV-VSVG 共同使用，使用方法见下文。 |
| 产品目录号: | #8449 |
| 稳定性: | 瞬表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
买家导航
pUMVC载体质粒基本信息
pUMVC质粒图谱载体图谱和pUMVC载体序列质粒序列多克隆位点信息
pUMVC质粒载体简介
双质粒系统逆病毒包装质粒pUMVC使用方法
Day 1
1. Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 mL of media. (This can be scaled up if desired)
Day 2
1. Set up (use polypropylene tubes for this; polystyrene tubes DO NOT work!):
1 µg retroviral DNA encoding gene X
1 µg packaging plasmid Mix
the packaging plasmid (pUMVC3) at a 8:1 ratio with the envelope plasmid (pCVM-VSV-G)-a total of 1µg
DME without serum to 94μL total
6 μL Fugene
Mix and wait 15 to 30 minutes at room temperature
Add to 293T cells without touching the sides of the dish (DO NOT CHANGE MEDIA)
If you are using amphotropic virus then move immediately to BL2+ in a secondary container, which has an absorbent material.
(This does not mean a couple of hours; it means Immediately!). The rest of this protocol is the same for all viruses---the BL2+ safety practices are in place if you are using amphotropic viruses
Day 3
1. Change the media to whatever media you wish to use when infecting target cells. 293T cells are easily detached so remember not to put the media directly onto to cells, but rather “run” it down the side of the dish. Remember that you will get the highest titer virus when your cells are “happy.”
2. Plate out your target cells
Day 4
1. Remove the medium from the 293T cells and use a 0.45 u syringe filter to remove any 293T cells. DO NOT use the 0.2 u filter, as it is likely to shear the envelope from your virus
making it noninfectious
Note: After filtering, the filter should be removed and placed in the biohazard bag in the hood and the syringe rinsed with bleach and decontaminated for a minimum of 20 min. It is useful to place a plastic beaker with bleach in the hood in advance
2. Add 8 to 10 μg/ml of polybrene (Hexadimethrine bromide) or protamine sulfate to the target cells
3. Carry out infection for 1 to 4 hours. Remove virus and replace with fresh media
Note: If you wish to do a second infection the following day, it is important to put fresh media on the cells and not let the virus remain on the cells overnight. The media contains huge amounts of envelope, both associated and unassociated with viral particles, which will bind all the cell surface receptors required for virus adsorption, resulting in their down-regulation. Hence, if you don’t change the media after the initial infection, very few receptors will be available for the next round of infection. In addition, very few cells tolerate the presence of high levels of the VSV-G envelope for extended periods of time (i.e. a lot of your cells may die)
Day 5 +
1. Allow the cells to recover and begin to express the virus-encoded genes. The cells usually require 48 hours for this to occur
2. Add drug if you are scoring for the presence of a vector that carries a drug resistance marker. Prior to this step it is advisable that you titrate the drug to be used for selection in order to know precisely how much to add. In addition, it is necessary to bring an extra plate of uninfected cells (often referred to as “canaries”) which will function as a positive control in the kill assay. Add drug to both plates. When your canaries are dead, you can remove the drug.
pUMVC质粒序列载体序列
LOCUS pUMVC Sequencing Result 9287 bp DNA SYN
DEFINITION pUMVC Sequencing Result
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..9287
/organism="pUMVC Sequencing Result"
/mol_type="other DNA"
gene complement(1246..2061)
/label="KanR2"
/gene="KanR2"
CDS complement(1246..2061)
/label="ORF frame 3"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
promoter complement(2207..2235)
/label="AmpR_promoter"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature complement(2394..2416)
/label="pGEX_3_primer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
promoter 2687..3263
/label="CMV_immearly_promoter"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature 2766..3053
/label="CAG_enhancer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature 3220..3240
/label="CMV_fwd_primer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
promoter 3221..3290
/label="CMV_promoter"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
promoter 3233..3352
/label="CMV2_promoter"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature 3264..3283
/label="pCEP_fwd_primer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature 3266..3290
/label="LNCX_primer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
CDS 3725..5341
/label="ORF frame 2"
/translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP
TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH
PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE
DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR
AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT
LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR
QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN
VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE
KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK
EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*"
misc_feature 3731..4138
/label="gag"
/translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP
TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH
PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE
DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR
AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT
LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR
QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN
VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE
KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK
EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*"
misc_feature 4074..4096
/label="MSCV_primer"
/translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP
TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH
PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE
DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR
AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT
LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR
QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN
VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE
KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK
EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*"
misc_feature 4074..4096
/label="pLXSN_5_primer"
/translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP
TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH
PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE
DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR
AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT
LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR
QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN
VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE
KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK
EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*"
misc_feature 4112..4128
/label="pBABE_5_primer"
/translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWP
TFNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVH
PKPPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTE
DPPPYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLR
AGGNGQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGT
LLTGEEKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYR
QLLLAGLQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETN
VSMSFIWQSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEE
KEERRRTEDEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCK
EKGHWAKDCPKKPRGPRGPRPQTSLLTLDD*"
CDS 5669..8941
/label="ORF frame 2"
/translation="MGPMGQPLQVLTLNIEDEYRLHETSKEPDVSLGSTWLSDFPQAW
AETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQ
SPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVL
DLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDL
ADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQ
VKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAP
LYPLTKTGTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVL
TQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHA
VEALVKQPPDRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLD
ILAEAHGTRPDLTDQPLPDADHTWYTDGSSLLQEGQRKAGAAVTTETEVIWAKALPAG
TSAQRAELIALTQALKMAEGKKLNVYTDSRYAFATAHIHGEIYRRRGLLTSEGKEIKN
KDEILALLKALFLPKRLSIIHCPGHQKGHSAEARGNRMADQAARKAAITETPDTSTLL
IENSSPYTSEHFHYTVTDIKDLTKLGAIYDKTKKYWVYQGKPVMPDQFTFELLDFLHQ
LTHLSFSKMKALLERSHSPYYMLNRDRTLKNITETCKACAQVNASKSAVKQGTRVRGH
RPGTHWEIDFTEIKPGLYGYKYLLVFIDTFSGWIEAFPTKKETAKVVTKKLLEEIFPR
FGMPQVLGTDNGPAFVSKVSQTVADLLGIDWKLHCAYRPQSSGQVERMNRTIKETLTK
LTLATGSRDWVLLLPLALYRARNTPGPHGLTPYEILYGAPPPLVNFPDPDMTRVTNSP
SLQAHLQALYLVQHEVWRPLAAAYQEQLDRPVVPHPYRVGDTVWVRRHQTKNLEPRWK
GPYPVLLTTPTALKVDGIAAWIHAAHVKAADPGGGPSSRLTWRVQRSQNPLKIRLTRE
AP*"
misc_feature 8816..8832
/label="pBMN_5_primer"
/translation="MGPMGQPLQVLTLNIEDEYRLHETSKEPDVSLGSTWLSDFPQAW
AETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQ
SPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVL
DLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDL
ADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQ
VKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAP
LYPLTKTGTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVL
TQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHA
VEALVKQPPDRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLD
ILAEAHGTRPDLTDQPLPDADHTWYTDGSSLLQEGQRKAGAAVTTETEVIWAKALPAG
TSAQRAELIALTQALKMAEGKKLNVYTDSRYAFATAHIHGEIYRRRGLLTSEGKEIKN
KDEILALLKALFLPKRLSIIHCPGHQKGHSAEARGNRMADQAARKAAITETPDTSTLL
IENSSPYTSEHFHYTVTDIKDLTKLGAIYDKTKKYWVYQGKPVMPDQFTFELLDFLHQ
LTHLSFSKMKALLERSHSPYYMLNRDRTLKNITETCKACAQVNASKSAVKQGTRVRGH
RPGTHWEIDFTEIKPGLYGYKYLLVFIDTFSGWIEAFPTKKETAKVVTKKLLEEIFPR
FGMPQVLGTDNGPAFVSKVSQTVADLLGIDWKLHCAYRPQSSGQVERMNRTIKETLTK
LTLATGSRDWVLLLPLALYRARNTPGPHGLTPYEILYGAPPPLVNFPDPDMTRVTNSP
SLQAHLQALYLVQHEVWRPLAAAYQEQLDRPVVPHPYRVGDTVWVRRHQTKNLEPRWK
GPYPVLLTTPTALKVDGIAAWIHAAHVKAADPGGGPSSRLTWRVQRSQNPLKIRLTRE
AP*"
ORIGIN
1 CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG
61 TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC
121 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA
181 AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT
241 CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG
301 GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG
361 CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT
421 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC
481 AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC
541 TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC
601 GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT
661 TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC
721 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG
781 AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA
841 ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA
901 CCTATCTCAG CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCGG GGGGGGGGGC
961 GCTGAGGTCT GCCTCGTGAA GAAGGTGTTG CTGACTCATA CCAGGCCTGA ATCGCCCCAT
1021 CATCCAGCCA GAAAGTGAGG GAGCCACGGT TGATGAGAGC TTTGTTGTAG GTGGACCAGT
1081 TGGTGATTTT GAACTTTTGC TTTGCCACGG AACGGTCTGC GTTGTCGGGA AGATGCGTGA
1141 TCTGATCCTT CAACTCAGCA AAAGTTCGAT TTATTCAACA AAGCCGCCGT CCCGTCAAGT
1201 CAGCGTAATG CTCTGCCAGT GTTACAACCA ATTAACCAAT TCTGATTAGA AAAACTCATC
1261 GAGCATCAAA TGAAACTGCA ATTTATTCAT ATCAGGATTA TCAATACCAT ATTTTTGAAA
1321 AAGCCGTTTC TGTAATGAAG GAGAAAACTC ACCGAGGCAG TTCCATAGGA TGGCAAGATC
1381 CTGGTATCGG TCTGCGATTC CGACTCGTCC AACATCAATA CAACCTATTA ATTTCCCCTC
1441 GTCAAAAATA AGGTTATCAA GTGAGAAATC ACCATGAGTG ACGACTGAAT CCGGTGAGAA
1501 TGGCAAAAGC TTATGCATTT CTTTCCAGAC TTGTTCAACA GGCCAGCCAT TACGCTCGTC
1561 ATCAAAATCA CTCGCATCAA CCAAACCGTT ATTCATTCGT GATTGCGCCT GAGCGAGACG
1621 AAATACGCGA TCGCTGTTAA AAGGACAATT ACAAACAGGA ATCGAATGCA ACCGGCGCAG
1681 GAACACTGCC AGCGCATCAA CAATATTTTC ACCTGAATCA GGATATTCTT CTAATACCTG
1741 GAATGCTGTT TTCCCGGGGA TCGCAGTGGT GAGTAACCAT GCATCATCAG GAGTACGGAT
1801 AAAATGCTTG ATGGTCGGAA GAGGCATAAA TTCCGTCAGC CAGTTTAGTC TGACCATCTC
1861 ATCTGTAACA TCATTGGCAA CGCTACCTTT GCCATGTTTC AGAAACAACT CTGGCGCATC
1921 GGGCTTCCCA TACAATCGAT AGATTGTCGC ACCTGATTGC CCGACATTAT CGCGAGCCCA
1981 TTTATACCCA TATAAATCAG CATCCATGTT GGAATTTAAT CGCGGCCTCG AGCAAGACGT
2041 TTCCCGTTGA ATATGGCTCA TAACACCCCT TGTATTACTG TTTATGTAAG CAGACAGTTT
2101 TATTGTTCAT GATGATATAT TTTTATCTTG TGCAATGTAA CATCAGAGAT TTTGAGACAC
2161 AACGTGGCTT TCCCCCCCCC CCATTATTGA AGCATTTATC AGGGTTATTG TCTCATGAGC
2221 GGATACATAT TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG CACATTTCCC
2281 CGAAAAGTGC CACCTGACGT CTAAGAAACC ATTATTATCA TGACATTAAC CTATAAAAAT
2341 AGGCGTATCA CGAGGCCCTT TCGTCTCGCG CGTTTCGGTG ATGACGGTGA AAACCTCTGA
2401 CACATGCAGC TCCCGGAGAC GGTCACAGCT TGTCTGTAAG CGGATGCCGG GAGCAGACAA
2461 GCCCGTCAGG GCGCGTCAGC GGGTGTTGGC GGGTGTCGGG GCTGGCTTAA CTATGCGGCA
2521 TCAGAGCAGA TTGTACTGAG AGTGCACCAT ATGCGGTGTG AAATACCGCA CAGATGCGTA
2581 AGGAGAAAAT ACCGCATCAG ATTGGCTATT GGCCATTGCA TACGTTGTAT CCATATCATA
2641 ATATGTACAT TTATATTGGC TCATGTCCAA CATTACCGCC ATGTTGACAT TGATTATTGA
2701 CTAGTTATTA ATAGTAATCA ATTACGGGGT CATTAGTTCA TAGCCCATAT ATGGAGTTCC
2761 GCGTTACATA ACTTACGGTA AATGGCCCGC CTGGCTGACC GCCCAACGAC CCCCGCCCAT
2821 TGACGTCAAT AATGACGTAT GTTCCCATAG TAACGCCAAT AGGGACTTTC CATTGACGTC
2881 AATGGGTGGA GTATTTACGG TAAACTGCCC ACTTGGCAGT ACATCAAGTG TATCATATGC
2941 CAAGTACGCC CCCTATTGAC GTCAATGACG GTAAATGGCC CGCCTGGCAT TATGCCCAGT
3001 ACATGACCTT ATGGGACTTT CCTACTTGGC AGTACATCTA CGTATTAGTC ATCGCTATTA
3061 CCATGGTGAT GCGGTTTTGG CAGTACATCA ATGGGCGTGG ATAGCGGTTT GACTCACGGG
3121 GATTTCCAAG TCTCCACCCC ATTGACGTCA ATGGGAGTTT GTTTTGGCAC CAAAATCAAC
3181 GGGACTTTCC AAAATGTCGT AACAACTCCG CCCCATTGAC GCAAATGGGC GGTAGGCGTG
3241 TACGGTGGGA GGTCTATATA AGCAGAGCTC GTTTAGTGAA CCGTCAGATC GCCTGGAGAC
3301 GCCATCCACG CTGTTTTGAC CTCCATAGAA GACACCGGGA CCGATCCAGC CTCCGCGGCC
3361 GGGAACGGTG CATTGGAACG CGGATTCCCC GTGCCAAGAG TGACGTAAGT ACCGCCTATA
3421 GACTCTATAG GCACACCCCT TTGGCTCTTA TGCATGCTAT ACTGTTTTTG GCTTGGGGCC
3481 TATACACCCC CGCTTCCTTA TGCTATAGGT GATGGTATAG CTTAGCCTAT AGGTGTGGGT
3541 TATTGACCAT TATTGACCAC TCCAACGGTG GAGGGCAGTG TAGTCTGAGC AGTACTCGTT
3601 GCTGCCGCGC GCGCCACCAG ACATAATAGC TGACAGACTA ACAGACTGTT CCTTTCCATG
3661 GGTCTTTTCT GCAGTCACCG TCGTCGACGG TATCGATAAG CTTGATATCG AATTCGCCGC
3721 CACCATGGGC CAGACTGTTA CCACTCCCTT AAGTTTGACC TTAGGTCACT GGAAAGATGT
3781 CGAGCGGATC GCTCACAACC AGTCGGTAGA TGTCAAGAAG AGACGTTGGG TTACCTTCTG
3841 CTCTGCAGAA TGGCCAACCT TTAACGTCGG ATGGCCGCGA GACGGCACCT TTAACCGAGA
3901 CCTCATCACC CAGGTTAAGA TCAAGGTCTT TTCACCTGGC CCGCATGGAC ACCCAGACCA
3961 GGTCCCCTAC ATCGTGACCT GGGAAGCCTT GGCTTTTGAC CCCCCTCCCT GGGTCAAGCC
4021 CTTTGTACAC CCTAAGCCTC CGCCTCCTCT TCCTCCATCC GCCCCGTCTC TCCCCCTTGA
4081 ACCTCCTCGT TCGACCCCGC CTCGATCCTC CCTTTATCCA GCCCTCACTC CTTCTCTAGG
4141 CGCCAAACCT AAACCTCAAG TTCTTTCTGA CAGTGGGGGG CCGCTCATCG ACCTACTTAC
4201 AGAAGACCCC CCGCCTTATA GGGACCCAAG ACCACCCCCT TCCGACAGGG ACGGAAATGG
4261 TGGAGAAGCG ACCCCTGCGG GAGAGGCACC GGACCCCTCC CCAATGGCAT CTCGCCTACG
4321 TGGGAGACGG GAGCCCCCTG TGGCCGACTC CACTACCTCG CAGGCATTCC CCCTCCGCGC
4381 AGGAGGAAAC GGACAGCTTC AATACTGGCC GTTCTCCTCT TCTGACCTTT ACAACTGGAA
4441 AAATAATAAC CCTTCTTTTT CTGAAGATCC AGGTAAACTG ACAGCTCTGA TCGAGTCTGT
4501 CCTCATCACC CATCAGCCCA CCTGGGACGA CTGTCAGCAG CTGTTGGGGA CTCTGCTGAC
4561 CGGAGAAGAA AAACAACGGG TGCTCTTAGA GGCTAGAAAG GCGGTGCGGG GCGATGATGG
4621 GCGCCCCACT CAACTGCCCA ATGAAGTCGA TGCCGCTTTT CCCCTCGAGC GCCCAGACTG
4681 GGATTACACC ACCCAGGCAG GTAGGAACCA CCTAGTCCAC TATCGCCAGT TGCTCCTAGC
4741 GGGTCTCCAA AACGCGGGCA GAAGCCCCAC CAATTTGGCC AAGGTAAAAG GAATAACACA
4801 AGGGCCCAAT GAGTCTCCCT CGGCCTTCCT AGAGAGACTT AAGGAAGCCT ATCGCAGGTA
4861 CACTCCTTAT GACCCTGAGG ACCCAGGGCA AGAAACTAAT GTGTCTATGT CTTTCATTTG
4921 GCAGTCTGCC CCAGACATTG GGAGAAAGTT AGAGAGGTTA GAAGATTTAA AAAACAAGAC
4981 GCTTGGAGAT TTGGTTAGAG AGGCAGAAAA GATCTTTAAT AAACGAGAAA CCCCGGAAGA
5041 AAGAGAGGAA CGTATCAGGA GAGAAACAGA GGAAAAAGAA GAACGCCGTA GGACAGAGGA
5101 TGAGCAGAAA GAGAAAGAAA GAGATCGTAG GAGACATAGA GAGATGAGCA AGCTATTGGC
5161 CACTGTCGTT AGTGGACAGA AACAGGATAG ACAGGGAGGA GAACGAAGGA GGTCCCAACT
5221 CGATCGCGAC CAGTGTGCCT ACTGCAAAGA AAAGGGGCAC TGGGCTAAAG ATTGTCCCAA
5281 GAAACCACGA GGACCTCGGG GACCAAGACC CCAGACCTCC CTCCTGACCC TAGATGACTA
5341 GGGAGGTCAG GGTCAGGAGC CCCCCCCTGA ACCCAGGATA ACCCTCAAAG TCGGGGGGCA
5401 ACCCGTCACC TTCCTGGTAG ATACTGGGGC CCAACACTCC GTGCTGACCC AAAATCCTGG
5461 ACCCCTAAGT GATAAGTCTG CCTGGGTCCA AGGGGCTACT GGAGGAAAGC GGTATCGCTG
5521 GACCACGGAT CGCAAAGTAC ATCTAGCTAC CGGTAAGGTC ACCCACTCTT TCCTCCATGT
5581 ACCAGACTGT CCCTATCCTC TGTTAGGAAG AGATTTGCTG ACTAAACTAA AAGCCCAAAT
5641 CCACTTTGAG GGATCAGGAG CTCAGGTTAT GGGACCAATG GGGCAGCCCC TGCAAGTGTT
5701 GACCCTAAAT ATAGAAGATG AGTATCGGCT ACATGAGACC TCAAAAGAGC CAGATGTTTC
5761 TCTAGGGTCC ACATGGCTGT CTGATTTTCC TCAGGCCTGG GCGGAAACCG GGGGCATGGG
5821 ACTGGCAGTT CGCCAAGCTC CTCTGATCAT ACCTCTGAAA GCAACCTCTA CCCCCGTGTC
5881 CATAAAACAA TACCCCATGT CACAAGAAGC CAGACTGGGG ATCAAGCCCC ACATACAGAG
5941 ACTGTTGGAC CAGGGAATAC TGGTACCCTG CCAGTCCCCC TGGAACACGC CCCTGCTACC
6001 CGTTAAGAAA CCAGGGACTA ATGATTATAG GCCTGTCCAG GATCTGAGAG AAGTCAACAA
6061 GCGGGTGGAA GACATCCACC CCACCGTGCC CAACCCTTAC AACCTCTTGA GCGGGCTCCC
6121 ACCGTCCCAC CAGTGGTACA CTGTGCTTGA TTTAAAGGAT GCCTTTTTCT GCCTGAGACT
6181 CCACCCCACC AGTCAGCCTC TCTTCGCCTT TGAGTGGAGA GATCCAGAGA TGGGAATCTC
6241 AGGACAATTG ACCTGGACCA GACTCCCACA GGGTTTCAAA AACAGTCCCA CCCTGTTTGA
6301 TGAGGCACTG CACAGAGACC TAGCAGACTT CCGGATCCAG CACCCAGACT TGATCCTGCT
6361 ACAGTACGTG GATGACTTAC TGCTGGCCGC CACTTCTGAG CTAGACTGCC AACAAGGTAC
6421 TCGGGCCCTG TTACAAACCC TAGGGAACCT CGGGTATCGG GCCTCGGCCA AGAAAGCCCA
6481 AATTTGCCAG AAACAGGTCA AGTATCTGGG GTATCTTCTA AAAGAGGGTC AGAGATGGCT
6541 GACTGAGGCC AGAAAAGAGA CTGTGATGGG GCAGCCTACT CCGAAGACCC CTCGACAACT
6601 AAGGGAGTTC CTAGGGACGG CAGGCTTCTG TCGCCTCTGG ATCCCTGGGT TTGCAGAAAT
6661 GGCAGCCCCC TTGTACCCTC TCACCAAAAC GGGGACTCTG TTTAATTGGG GCCCAGACCA
6721 ACAAAAGGCC TATCAAGAAA TCAAGCAAGC TCTTCTAACT GCCCCAGCCC TGGGGTTGCC
6781 AGATTTGACT AAGCCCTTTG AACTCTTTGT CGACGAGAAG CAGGGCTACG CCAAAGGTGT
6841 CCTAACGCAA AAACTGGGAC CTTGGCGTCG GCCGGTGGCC TACCTGTCCA AAAAGCTAGA
6901 CCCAGTAGCA GCTGGGTGGC CCCCTTGCCT ACGGATGGTA GCAGCCATTG CCGTACTGAC
6961 AAAGGATGCA GGCAAGCTAA CCATGGGACA GCCACTAGTC ATTCTGGCCC CCCATGCAGT
7021 AGAGGCACTA GTCAAACAAC CCCCCGACCG CTGGCTTTCC AACGCCCGGA TGACTCACTA
7081 TCAGGCCTTG CTTTTGGACA CGGACCGGGT CCAGTTCGGA CCGGTGGTAG CCCTGAACCC
7141 GGCTACGCTG CTCCCACTGC CTGAGGAAGG GCTGCAACAC AACTGCCTTG ATATCCTGGC
7201 CGAAGCCCAC GGAACCCGAC CCGACCTAAC GGACCAGCCG CTCCCAGACG CCGACCACAC
7261 CTGGTACACG GATGGAAGCA GTCTCTTACA AGAGGGACAG CGTAAGGCGG GAGCTGCGGT
7321 GACCACCGAG ACCGAGGTAA TCTGGGCTAA AGCCCTGCCA GCCGGGACAT CCGCTCAGCG
7381 GGCTGAACTG ATAGCACTCA CCCAGGCCCT AAAGATGGCA GAAGGTAAGA AGCTAAATGT
7441 TTATACTGAT AGCCGTTATG CTTTTGCTAC TGCCCATATC CATGGAGAAA TATACAGAAG
7501 GCGTGGGTTG CTCACATCAG AAGGCAAAGA GATCAAAAAT AAAGACGAGA TCTTGGCCCT
7561 ACTAAAAGCC CTCTTTCTGC CCAAAAGACT TAGCATAATC CATTGTCCAG GACATCAAAA
7621 GGGACACAGC GCCGAGGCTA GAGGCAACCG GATGGCTGAC CAAGCGGCCC GAAAGGCAGC
7681 CATCACAGAG ACTCCAGACA CCTCTACCCT CCTCATAGAA AATTCATCAC CCTACACCTC
7741 AGAACATTTT CATTACACAG TGACTGATAT AAAGGACCTA ACCAAGTTGG GGGCCATTTA
7801 TGATAAAACA AAGAAGTATT GGGTCTACCA AGGAAAACCT GTGATGCCTG ACCAGTTTAC
7861 TTTTGAATTA TTAGACTTTC TTCATCAGCT GACTCACCTC AGCTTCTCAA AAATGAAGGC
7921 TCTCCTAGAG AGAAGCCACA GTCCCTACTA CATGCTGAAC CGGGATCGAA CACTCAAAAA
7981 TATCACTGAG ACCTGCAAAG CTTGTGCACA AGTCAACGCC AGCAAGTCTG CCGTTAAACA
8041 GGGAACTAGG GTCCGCGGGC ATCGGCCCGG CACTCATTGG GAGATCGATT TCACCGAGAT
8101 AAAGCCCGGA TTGTATGGCT ATAAATATCT TCTAGTTTTT ATAGATACCT TTTCTGGCTG
8161 GATAGAAGCC TTCCCAACCA AGAAAGAAAC CGCCAAGGTC GTAACCAAGA AGCTACTAGA
8221 GGAGATCTTC CCCAGGTTCG GCATGCCTCA GGTATTGGGA ACTGACAATG GGCCTGCCTT
8281 CGTCTCCAAG GTGAGTCAGA CAGTGGCCGA TCTGTTGGGG ATTGATTGGA AATTACATTG
8341 TGCATACAGA CCCCAAAGCT CAGGCCAGGT AGAAAGAATG AATAGAACCA TCAAGGAGAC
8401 TTTAACTAAA TTAACGCTTG CAACTGGCTC TAGAGACTGG GTGCTCCTAC TCCCCTTAGC
8461 CCTGTACCGA GCCCGCAACA CGCCGGGCCC CCATGGCCTC ACCCCATATG AGATCTTATA
8521 TGGGGCACCC CCGCCCCTTG TAAACTTCCC TGACCCTGAC ATGACAAGAG TTACTAACAG
8581 CCCCTCTCTC CAAGCTCACT TACAGGCTCT CTACTTAGTC CAGCACGAAG TCTGGAGACC
8641 TCTGGCGGCA GCCTACCAAG AACAACTGGA CCGACCGGTG GTACCTCACC CTTACCGAGT
8701 CGGCGACACA GTGTGGGTCC GCCGACACCA GACTAAGAAC CTAGAACCTC GCTGGAAAGG
8761 ACCTTACCCA GTCCTGCTGA CCACCCCCAC CGCCCTCAAA GTAGACGGCA TCGCAGCTTG
8821 GATACACGCC GCCCACGTGA AGGCTGCCGA CCCCGGGGGT GGACCATCCT CTAGACTGAC
8881 ATGGCGCGTT CAACGCTCTC AAAACCCCTT AAAAATAAGG TTAACCCGCG AGGCCCCCTA
8941 ATCCCCTTAA TTCTTCTGAT GCTCAGAGGG GTCAGTGTGG GCCCGGTACC GTATACTCTA
9001 GAGCGGCCGC GGATCCAGAT CTTTTTCCCT CTGCCAAAAA TTATGGGGAC ATCATGAAGC
9061 CCCTTGAGCA TCTGACTTCT GGCTAATAAA GGAAATTTAT TTTCATTGCA ATAGTGTGTT
9121 GGAATTTTTT GTGTCTCTCA CTCGGAAGGA CATATGGGAG GGCAAATCAT TTAAAACATC
9181 AGAATGAGTA TTTGGTTTAG AGTTTGGCAA CATATGCCCA TTCTTCCGCT TCCTCgctca
9241 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagct
//
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pUMVC质粒载体应用举例
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