| 质粒名称: | pCAGGs-RFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 127347 |
| 存储实验室: | Phil Sharp |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGs |
| 载体出品公司: | Allan Gurtan (Phil Sharp Lab) |
| 空载体大小: | 5438bp |
| 完整质粒大小: | 5438bp |
| 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Red Fluorescent Protein、RFP |
| 物种来源: | Synthetic |
| 插入大小: | 714 bp |
| 启动子: | CAGGS Promoter |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | GGTTCGGCTTCTGGCGTGTGACC |
| 3’测序引物 | CCCATATGTCCTTCCGAGTG |
|
文献引用方法 |
|
| 材料和方法部分: | pCAGGs-RFP was a gift from Phil Sharp (Addgene plasmid # 127347 ; http://n2t.net/addgene:127347 ; RRID:Addgene_127347) |
买家导航
pCAGGs-RFP载体质粒基本信息
pCAGGs-RFP质粒图谱载体图谱和pCAGGs-RFP载体序列质粒序列多克隆位点信息
pCAGGs-RFP质粒载体简介
pCAGGs-RFP质粒序列载体序列
LOCUS pCAGGs-RFP 5438 bp DNA circular SYN 15-DEC-2025
DEFINITION Expresses RFP constitutively via the CAGGS promoter in mammalian
cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5438)
TITLE Constitutive Fluorecent Protein Expression in Mammalian Cells via
the pCAGGs Promoter
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5438)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT Sequence Label: pCAGGs-RFP
FEATURES Location/Qualifiers
source 1..5438
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 385..662
/label=chicken beta-actin promoter
intron 663..1679
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1603..1625
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1687..1706
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1733..1742
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1739..2452
/codon_start=1
/product="monomeric derivative of red fluorescent protein
from Entacmaea quadricolor (Merzlyak et al., 2007)"
/label=TagRFP
/note="mammalian codon-optimized"
/translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
ARYCDLPSKLGHKLN"
primer_bind complement(2459..2478)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2524..2579
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2578..2597)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2940..2956)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2940..2956)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2953..2975)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2964..2980
/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(join(2988..2994,2995..3012,3013..3018))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3033..3054
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3113..3308
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3159..3294
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 3314..3448
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3351..3370)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3405..3424
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3516..3533)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3687..4275)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3767..3786)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(4446..5237,5238..5306))
/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 5069..5088
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5307..5411)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggggcgcg cgccrggggg gggggggggg ggsggggggs
541 ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
1261 cggggccgcc tcgggccggg grgggctcgg gggrggggcg cggcggcccc cggagcgccg
1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgccaccat
1741 ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg
1801 caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
1861 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
1921 cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc agggcatccc
1981 cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
2041 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
2101 caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac
2161 actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc tggaaggcag
2221 aagcgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac
2281 atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacca
2341 cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc acgaggtggc
2401 tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatt gagctcgatc
2461 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
2521 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
2581 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
2641 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
2701 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
2761 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
2821 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
2881 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
2941 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
3001 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
3061 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
3121 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
3181 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
3241 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
3301 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
3361 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
3421 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
3481 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
3541 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
3601 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
3661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
3721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
3781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
3841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
3901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
3961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
4021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
4081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
4141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
4201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
4261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
4321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
4381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
4441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
4501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
4561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
4621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
4681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
4741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
4801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
4861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
4921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
4981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
5041 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
5101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
5161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
5221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
5281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
5341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
5401 ggggttccgc gcacatttcc ccgaaaagtg ccacctgg
//
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