| 质粒名称: | pCMV-T7-EGFP (BPK1098) |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 133962 |
| 存储实验室: | Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV-T7 |
| 空载体大小: | 4119 bp |
| 完整质粒大小: | 4119 bp |
| 载体修饰: | EGFP inserted into NotI/AgeI restriction sites |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | EGFP |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 717 bp |
| 启动子: | CMV and T7 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI(未破坏) |
| 3’克隆位点: | AgeI(未破坏) |
| 5’测序引物 | CMV forward - CGCAAATGGGCGGTAGGCGTG |
| 3’测序引物 | BGH reverse - TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-T7-EGFP (BPK1098) was a gift from Benjamin Kleinstiver (Addgene plasmid # 133962 ; http://n2t.net/addgene:133962 ; RRID:Addgene_133962) |
| 参考文献部分: | |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
133962-质粒图谱.png)
载体简介
载体序列
LOCUS pCMV-T7-EGFP (BPK1098) 4119 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CMV and T7 promoter plasmid for EGFP expression in mammalian cells
or for in vitro transcription.
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-EGFP (BPK1098)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4119)
TITLE CMV and T7 promoter plasmid to express EGFP (BPK1098)
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4119)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4119
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 132..335
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 286..306
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 377..396
/label=T7
/note="T7 promoter, forward primer"
promoter 377..395
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 403..412
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 409..1125
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(454..475)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(715..734)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1062..1083
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1134..1151
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1174..1191)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1180..1404
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(1475..1491)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1475..1491)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1488..1510)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1499..1515
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(1523..1553)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1568..1589
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1706..1723)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1877..2465)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1957..1976)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2636..3496)
/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 3259..3278
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3497..3601)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3680..3699)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3871..131
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggtgagcaag
421 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
481 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
541 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
601 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
661 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
721 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
781 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
841 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
901 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
961 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
1021 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
1081 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgacc ggtcatcatc
1141 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
1201 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
1261 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
1321 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
1381 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgatac
1441 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
1501 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg
1561 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
1621 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
1681 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
1741 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
1801 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
1861 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
1921 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
1981 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
2041 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
2101 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
2161 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
2221 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
2281 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
2341 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
2401 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
2461 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
2521 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
2581 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
2641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
2701 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
2761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
2821 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
2881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
2941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
3001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
3061 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
3121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
3181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
3241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
3301 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
3361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
3421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
3481 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
3541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
3601 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc
3661 ccctagggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
3721 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
3781 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
3841 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
3901 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
3961 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccaatgacg tcaataatga
4021 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
4081 tacggtaaac tgcccacttg gcagtacatc aagtgtatc
//
