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pGEM-3Zf(+)体外转录质粒 货号：P1015
来源:
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作者:shhebio
|
发布时间: 2017-11-28
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215 次浏览
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信息
| 启动子 | Lac/lac |
| 复制子 | pUC |
| 质粒分类 | 大肠杆菌载体；克隆质粒 |
| 质粒大小 | 3197bp |
| 质粒标签 | LacZ |
| 原核抗性 | Amp |
| 克隆菌株 | DH5α |
| 培养条件 | 37℃ |
| 5'测序引物 | M13R:CAGGAAACAGCTATGACC |
| 3'测序引物 | M13F:TGTAAAACGACGGCCAGT |
| 备注 | 蓝白斑筛选 |
| 质粒宿主 | 大肠杆菌 |
| 质粒用途 | 基因模板 |
| 片段类型 | 基因CDS |
| 片段物种 | 空载体 |
| 原核抗性 | 氨苄青霉素 |
质粒简介
pGEM-3Zf(+)可作为一般克隆载体和高效率RNA体外合成的模板，此载体含SP6和T7RNA聚合酶启动子，还含有lacZct-肽基因及单克隆位点，
可在含IPTG和X Gal的平板上对重组子进行蓝/白筛选。此载体含有丝状噬菌体fl的复制起点，可诱导产生单链DNA。pGEM 3Zf(+)表示其fl是正方向的，
pGEM-3Zf(+)特征图谱见图
_conew1.jpg)
1_conew1.jpg)
可在含IPTG和X Gal的平板上对重组子进行蓝/白筛选。此载体含有丝状噬菌体fl的复制起点，可诱导产生单链DNA。pGEM 3Zf(+)表示其fl是正方向的，
pGEM-3Zf(+)特征图谱见图
序列
LOCUS Exported 3197 bp ds-DNA circular SYN 24-JUN-2015
DEFINITION Vector for standard cloning, efficient in vitro RNA synthesis, and
ssDNA production. Identical to pGEM(R)?3Zf(-) except for the
orientation of the f1 origin.
ACCESSION X65306
VERSION .
KEYWORDS pGEM-3Zf(+)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3197)
AUTHORS Promega
TITLE Direct Submission
JOURNAL Exported Thursday, September 1, 2016 from SnapGene Viewer 3.1.4
http://www.shhebio.com
FEATURES Location/Qualifiers
source 1..3197
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
misc_feature 5..61
/note="MCS"
/note="pUC18/19 multiple cloning site"
promoter complement(68..86)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(104..120)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 128..144
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(152..182)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(506..1094)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1265..2125)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2126..2230)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(2562..3017)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2994..108)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
primer_bind 3158..3174
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3181..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gggcgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
61 tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag ctgtttcctg
121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
241 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
301 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
1021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
1081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
1141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
1201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
1261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
1321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
1381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
1441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
1501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
1561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
1621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
1681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
1741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
1801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
1861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
1921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
1981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
2041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
2101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
2161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
2221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
2281 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
2341 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2401 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
2461 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
2521 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
2581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
2641 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
2701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
2761 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
2821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
2881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
2941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
3001 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
3061 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
3121 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
3181 taatacgact cactata
//
DEFINITION Vector for standard cloning, efficient in vitro RNA synthesis, and
ssDNA production. Identical to pGEM(R)?3Zf(-) except for the
orientation of the f1 origin.
ACCESSION X65306
VERSION .
KEYWORDS pGEM-3Zf(+)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3197)
AUTHORS Promega
TITLE Direct Submission
JOURNAL Exported Thursday, September 1, 2016 from SnapGene Viewer 3.1.4
http://www.shhebio.com
FEATURES Location/Qualifiers
source 1..3197
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
misc_feature 5..61
/note="MCS"
/note="pUC18/19 multiple cloning site"
promoter complement(68..86)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(104..120)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 128..144
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(152..182)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(506..1094)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1265..2125)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2126..2230)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(2562..3017)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2994..108)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
primer_bind 3158..3174
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3181..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gggcgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
61 tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag ctgtttcctg
121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
241 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
301 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
1021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
1081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
1141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
1201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
1261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
1321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
1381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
1441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
1501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
1561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
1621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
1681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
1741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
1801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
1861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
1921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
1981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
2041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
2101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
2161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
2221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
2281 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
2341 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2401 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
2461 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
2521 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
2581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
2641 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
2701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
2761 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
2821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
2881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
2941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
3001 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
3061 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
3121 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
3181 taatacgact cactata
//
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标题摘要内容