| 出品公司: | CLONTECH |
|---|---|
| 载体名称: | pGFPuv |
| 质粒类型: | 大肠杆菌表达载体；基因载体 |
| 高拷贝/低拷贝: | 高 |
| 启动子: | lac |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 3337 bp |
| 5 测序引物及序列: | lac_promoter: CACTTTATGCTTCCGGCTCG |
| 3 测序引物及序列: | -- |
| 载体标签: | |
| 载体抗性: | Ampicillin |
| 筛选标记: | -- |
| 备注: | 含有GFPuv基因 |
| 产品目录号: | -- |
| 稳定性: | -- |
| 组成型: | -- |
| 病毒/非病毒: | -- |
英文名称： pGFPuv大肠杆菌载体
试剂级别： 分子生物学级
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
pGFPuv carries the“cycle 3”variant of GFP described by Crameri et al. (1). This gene was cloned between the two MCSs of the pUC19 derivative pPD16.43 (2). The GFPuv gene canbe easily excised from pGFPuv. Alternatively, the GFPuv coding sequence can be amplified by PCR. The GFPuv gene was inserted in frame with the lacZ initiation codon from pUC19 so that a β-galactosidase-GFPuv fusionprotein is expressed from the lac promoter in E. coli . Note, however, that if you excise the GFPuv coding sequence using a restriction site in the 5 MCS, the resulting fragment will encode the native (i.e.,non-fusion) GFPuv protein. The pUC backbone of pGFPuv provides a high copy number origin of replication and ampicillin resistance gene for propagation in E. coli.
pGFPuv carries the“cycle 3”variant of GFP described by Crameri et al. (1). This gene was cloned between the two MCSs of the pUC19 derivative pPD16.43 (2). The GFPuv gene canbe easily excised from pGFPuv. Alternatively, the GFPuv coding sequence can be amplified by PCR. The GFPuv gene was inserted in frame with the lacZ initiation codon from pUC19 so that a β-galactosidase-GFPuv fusionprotein is expressed from the lac promoter in E. coli . Note, however, that if you excise the GFPuv coding sequence using a restriction site in the 5 MCS, the resulting fragment will encode the native (i.e.,non-fusion) GFPuv protein. The pUC backbone of pGFPuv provides a high copy number origin of replication and ampicillin resistance gene for propagation in E. coli.
载体序列
LOCUS pGFPuv 3337 bp DNA SYNDEFINITION pGFPuvACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors.FEATURES Location/Qualifiers source 1..3337 /organism="pGFPuv" /mol_type="other DNA" promoter 143..172 /label="lac_promoter" misc_feature 186..208 /label="M13_pUC_rev_primer" promoter 207..225 /label="M13_reverse_primer" CDS 217..1005 /label="ORF frame 1" gene 295..990 /label="GFP (0 - 695)" /gene="GFP (0 - 695)" misc_feature complement(319..347) /label="GFP_R_primer" misc_feature complement(363..382) /label="GFPfusionREV_primer" misc_feature 942..962 /label="GFP_F_primer" misc_feature complement(1127..1149) /label="pGEX_3_primer" promoter 1467..1495 /label="AmpR_promoter" gene 1537..2397 /label="Ampicillin" /gene="Ampicillin" CDS 1537..2397 /label="ORF frame 1" rep_origin 2552..3171 /label="pBR322_origin"ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg 241 catgcctgca ggtcgactct agaggatccc cgggtaccgg tagaaaaaat gagtaaagga 301 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg 361 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt 421 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc 481 tcttatggtg ttcaatgctt ttcccgttat ccggatcata tgaaacggca tgactttttc 541 aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg 601 aactacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag 661 ttaaaaggta ttgattttaa agaagatgga aacattctcg gacacaaact cgagtacaac 721 tataactcac acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac 781 ttcaaaattc gccacaacat tgaagatgga tccgttcaac tagcagacca ttatcaacaa 841 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa 901 tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta 961 actgctgctg ggattacaca tggcatggat gagctctaca aataatgaat tccaactgag 1021 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggcct actagtcggc 1081 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 1141 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 1201 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 1261 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 1321 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc 1381 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 1441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 1501 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 1561 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 1621 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 1681 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 1741 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 1801 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 1861 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 1921 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 1981 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 2041 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 2101 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 2161 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 2221 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 2281 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 2341 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 2401 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 2461 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 2521 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 2581 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 2641 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 2701 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 2761 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 2821 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 2881 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 2941 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 3001 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 3061 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 3121 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 3181 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 3241 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 3301 accgagcgca gcgagtcagt gagcgaggaa gcggaag//
LOCUS pGFPuv 3337 bp DNA SYNDEFINITION pGFPuvACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors.FEATURES Location/Qualifiers source 1..3337 /organism="pGFPuv" /mol_type="other DNA" promoter 143..172 /label="lac_promoter" misc_feature 186..208 /label="M13_pUC_rev_primer" promoter 207..225 /label="M13_reverse_primer" CDS 217..1005 /label="ORF frame 1" gene 295..990 /label="GFP (0 - 695)" /gene="GFP (0 - 695)" misc_feature complement(319..347) /label="GFP_R_primer" misc_feature complement(363..382) /label="GFPfusionREV_primer" misc_feature 942..962 /label="GFP_F_primer" misc_feature complement(1127..1149) /label="pGEX_3_primer" promoter 1467..1495 /label="AmpR_promoter" gene 1537..2397 /label="Ampicillin" /gene="Ampicillin" CDS 1537..2397 /label="ORF frame 1" rep_origin 2552..3171 /label="pBR322_origin"ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg 241 catgcctgca ggtcgactct agaggatccc cgggtaccgg tagaaaaaat gagtaaagga 301 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg 361 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt 421 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc 481 tcttatggtg ttcaatgctt ttcccgttat ccggatcata tgaaacggca tgactttttc 541 aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg 601 aactacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag 661 ttaaaaggta ttgattttaa agaagatgga aacattctcg gacacaaact cgagtacaac 721 tataactcac acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac 781 ttcaaaattc gccacaacat tgaagatgga tccgttcaac tagcagacca ttatcaacaa 841 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa 901 tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta 961 actgctgctg ggattacaca tggcatggat gagctctaca aataatgaat tccaactgag 1021 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggcct actagtcggc 1081 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 1141 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 1201 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 1261 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 1321 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc 1381 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 1441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 1501 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 1561 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 1621 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 1681 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 1741 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 1801 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 1861 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 1921 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 1981 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 2041 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 2101 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 2161 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 2221 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 2281 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 2341 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 2401 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 2461 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 2521 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 2581 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 2641 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 2701 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 2761 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 2821 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 2881 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 2941 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 3001 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 3061 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 3121 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 3181 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 3241 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 3301 accgagcgca gcgagtcagt gagcgaggaa gcggaag//
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