| 质粒名称: | pCAG-mGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 14757 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6122 bp |
| 完整质粒大小: | 6953 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | membrane bound form of EGFP,GFP,green fluorescent protein |
| 物种来源: | -- |
| 插入大小: | 831 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | pCAG |
| 标签/融合蛋白: | Palmitoylation signal（插入件上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI (未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | pCAG-F |
| 3' 测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-mGFP was a gift from Connie Cepko (Addgene plasmid # 14757 ; http://n2t.net/addgene:14757 ; RRID:Addgene_14757) |
| 参考文献部分: | Controlled expression of transgenes introduced by in vivo electroporation. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2007 Jan 5. ():. 10.1073/pnas.0610155104 PubMed 17209010 |
买家导航
pCAGmGFP载体基本信息
pCAGmGFP载体图谱质粒图谱和多克隆位点信息
pCAGmGFP载体简介
pCAGmGFP载体序列
LOCUS pCAG-mGFP 6957 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCAG-mGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6957)
AUTHORS Matsuda T, Cepko CL
TITLE Controlled expression of transgenes introduced by in vivo
electroporation.
JOURNAL Proc Natl Acad Sci U S A. 2007 Jan 5. ():.
PUBMED 17209010
REFERENCE 2 (bases 1 to 6957)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6957
/organism="synthetic DNA construct"
/mol_type="other DNA"
intron 337..1353
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1277..1299
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1361..1380
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1495..2214
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1540..1561)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1801..1820)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2148..2169
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2317..2336)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2382..2437
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2436..2455)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2798..2814)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2798..2814)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2811..2833)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2822..2838
/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(2846..2876)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2891..2912
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2971..3166
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3017..3152
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3079..3098
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3172..3306
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3209..3228)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3263..3282
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3374..3391)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3545..4133)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3625..3644)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5645..6502)
/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
LIKH"
primer_bind 6265..6284
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6503..6607)
/gene="bla"
/label=AmpR promoter
enhancer 6638..60
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
61 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
121 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
181 ggsgcgccgg gggggggggg gggggggggg gggggggggg gggsgrggsg grgrggkgcg
241 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
301 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
361 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
421 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
481 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
541 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
601 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
661 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
721 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
781 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
841 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
901 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggrgggc
961 tcgggggrgg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
1021 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
1081 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
1141 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
1201 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
1261 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
1321 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
1381 ctgtctcatc attttggcaa agaattcctc gaggatgatc caccggtcgc caccatgctg
1441 tgctgtatga gaagaaccaa acaggttgaa aagaatgatg aggaccaaaa gatcatggtg
1501 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
1561 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
1621 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
1681 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
1741 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
1801 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
1861 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
1921 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
1981 aaggcgaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
2041 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
2101 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
2161 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
2221 ccggaattat cgcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
2281 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
2341 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
2401 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
2461 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
2521 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
2581 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
2641 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
2701 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
2761 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
2821 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
2881 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
2941 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
3001 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
3061 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
3121 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
3181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
3241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
3301 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
3361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
3421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
3481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
3541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
3601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
3661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
3721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
3781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
3841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
3901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
3961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
4021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
4081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
4141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
4201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
4261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatcc
4321 tgatgaatcc cctaatgatt tttatcaaaa tcattaagtt aaggtagata cacatcttgt
4381 catatgatca aatggtttcg ccaaaaatca ataatcagac aacaaaatgt gcgaactcga
4441 tattttacac gactctcttt accaattctg ccccgaatta cacttaaaac gactcaacag
4501 cttaacgttg gcttgccacg ccttacttga ctgtaaaact ctcactctta ccgaacttgg
4561 ccgtaacctg ccaaccaaag cgagaacaaa acataacatc aaacgaatcg accgattgtt
4621 aggtaatcgt cacctccaca aagagcgact cgctgtatac cgttggcatg ctagctttat
4681 ctgttcgggc aatacgatgc ccattgtact tgttgactgg tctgatatcc gtgagcaaaa
4741 acggcttatg gtattgcgag cttcagtcgc actacacggt cgttctgtta ctctttatga
4801 gaaagcgttc ccgctttcag agcaatgttc aaagaaagct catgaccaat ttctagccga
4861 ccttgcgagc attctaccga gtaacaccac accgctcatt gtcagtgatg ctggctttaa
4921 agtgccatgg tataaatccg ttgagaagct gggttggtac tggttaagtc gagtaagagg
4981 aaaagtacaa tatgcagacc taggagcgga aaactggaaa cctatcagca acttacatga
5041 tatgtcatct agtcactcaa agactttagg ctataagagg ctgactaaaa gcaatccaat
5101 ctcatgccaa attctattgt ataaatctcg ctctaaaggc cgaaaaaatc agcgctcgac
5161 acggactcat tgtcaccacc cgtcacctaa aatctactca gcgtcggcaa aggagccatg
5221 gattctagca actaacttac ctgttgaaat tcgaacaccc aaacaacttg ttaatatcta
5281 ttcgaagcga atgcagattg aagaaacctt ccgagacttg aaaagtcctg cctacggact
5341 aggcctacgc catagccgaa cgagcagctc agagcgtttt gatatcatgc tgctaatcgc
5401 cctgatgctt caactaacat gttggcttgc gggcgttcat gctcagaaac aaggttggga
5461 caagcacttc caggctaaca cagtcagaaa tcgaaacgta ctctcaacag ttcgcttagg
5521 catggaagtt ttgcggcatt ctggctacac aataacaagg gaagacttac tcgtggctgc
5581 aaccctacta gctcaaaatt tattcacaca tggttacgct ttggggaaat tatgagggga
5641 tctctcagtg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
5701 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
5761 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
5821 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
5881 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
5941 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
6001 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
6061 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
6121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
6181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
6241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
6301 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
6361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
6421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
6481 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
6541 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
6601 ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt gactagttat
6661 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
6721 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
6781 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
6841 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
6901 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatg
//
