| 质粒名称: | pRetroX-Tight-MCS_PGK-GpNLuc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 70185 |
| 存储实验室: | Antonio Amelio |
|
载体骨架基本信息 |
|
| 载体名称: | pRetroX-Tight-Pur |
| 载体出品公司: | Clontech |
| 空载体大小: | 7150bp |
| 完整质粒大小: | 7144bp |
| 载体类型: | 哺乳细胞表达, Retroviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 启动子: | PGK |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CTGGGCTCAGAGGCTGGG |
| 3’测序引物 | CCTACAGGTGGGGTCTTTC |
|
文献引用方法 |
|
| 材料和方法部分: | pRetroX-Tight-MCS_PGK-GpNLuc was a gift from Antonio Amelio (Addgene plasmid # 70185 ; http://n2t.net/addgene:70185 ; RRID:Addgene_70185) |
| 参考文献部分: | Fluorophore-NanoLuc BRET Reporters Enable Sensitive In Vivo Optical Imaging and Flow Cytometry for Monitoring Tumorigenesis. Schaub FX, Reza MS, Flaveny CA, Li W, Musicant AM, Hoxha S, Guo M, Cleveland JL, Amelio AL. Cancer Res. 2015 Sep 30. pii: canres.3538.2014. 10.1158/0008-5472.CAN-14-3538 PubMed 26424696 |
买家导航
pRetroX-Tight-MCS_PGK-GpNLuc载体质粒基本信息
pRetroX-Tight-MCS_PGK-GpNLuc质粒图谱载体图谱和pRetroX-Tight-MCS_PGK-GpNLuc载体序列质粒序列多克隆位点信息
pRetroX-Tight-MCS_PGK-GpNLuc质粒载体简介
pRetroX-Tight-MCS_PGK-GpNLuc质粒序列载体序列
LOCUS pRetroX-Tight-MCS_PGK-GpNLuc 7144 bp DNA circular SYN 14-APR-2025
DEFINITION Constiutive mammalian expression of the bioluminescent GpNLuc
LumiFluor reporter for single-cell analysis and sensitive optical
imaging of living subjects...
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7144)
AUTHORS Schaub FX, Reza MS, Flaveny CA, Li W, Musicant AM, Hoxha S, Guo M,
Cleveland JL, Amelio AL
TITLE Fluorophore-NanoLuc BRET Reporters Enable Sensitive In Vivo Optical
Imaging and Flow Cytometry for Monitoring Tumorigenesis.
JOURNAL Cancer Res. 2015 Sep 30. pii: canres.3538.2014.
PUBMED 26424696
REFERENCE 2 (bases 1 to 7144)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cancer Res.
2015 Sep 30. pii: canres.3538.2014."
COMMENT Sequence Label: pRetroX-Tight-MCS_PGK-GpNLuc
FEATURES Location/Qualifiers
source 1..7144
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 310..513
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 514..689
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 752..951
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1152..1568
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1498..1520
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1536..1552
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
promoter 1604..1918
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 1612..1630
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1648..1666
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1683..1701
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1719..1737
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1755..1773
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1790..1808
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1826..1844
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 1891..1915
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1974..2473
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(2009..2032)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 2406..2425
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS join(2488..2490,2491..2493,2494..3204)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(2488..2490,2491..2493,2494..3204)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2533..2554)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2794..2813)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3141..3162
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3220..3735
/codon_start=1
/product="NanoLuc(R) luciferase"
/label=Nluc
/note="human codon-optimized"
/translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
RLCERILA"
LTR 3850..4275
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
primer_bind complement(4378..4400)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4536..4556)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4541..4870
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4721..4856
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4892..4911
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4992..5009)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5163..5751)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5243..5262)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5922..6713,6714..6782))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6545..6564
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6783..6887)
/gene="bla"
/label=AmpR promoter
primer_bind 6955..6973
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
enhancer join(7073..7144,1..309)
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgctctg gcattatgcc
241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
481 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact
541 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
601 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
661 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
721 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
781 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
841 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
901 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
961 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
1021 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
1081 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
1141 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
1201 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
1261 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
1321 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
1381 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
1441 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
1501 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
1561 taggcgccgg aattgaagat ctgaggccct ttcgtcttca ctcgagttta ctccctatca
1621 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
1681 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
1741 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
1801 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
1861 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
1921 gagaaggatc cgcggccgcg ccggctctag atcgcgaacg cgtgaattct accgggtagg
1981 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
2041 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
2101 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
2161 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
2221 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
2281 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
2341 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
2401 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
2461 tccgggcctt tcgacctgca gcccaagatg gtgagcaagg gcgaggagct gttcaccggg
2521 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
2581 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
2641 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
2701 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
2761 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
2821 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
2881 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
2941 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
3001 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
3061 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
3121 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
3181 ctcggcatgg acgagctgta caaggatatc tccggaggta tggtcttcac actcgaagat
3241 ttcgttgggg actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga
3301 ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc
3361 ctgagcggtg aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg
3421 agcggcgacc aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat
3481 catcacttta aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac
3541 atgatcgact atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc
3601 actgtaacag ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc
3661 gacggctccc tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa
3721 cgcattctgg cgtaaatcga tagtggtcca ggctctagtt ttgactcaac aatatcacca
3781 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
3841 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
3901 ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac
3961 agatggaaca gggtcgaccc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
4021 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
4081 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
4141 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
4201 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
4261 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
4321 acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
4381 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
4441 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
4501 gtcacgtagc gatagcggag tgtagatccg gctgtggaat gtgtgtcagt tagggtgtgg
4561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
4621 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
4681 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
4741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
4801 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
4861 cttttgcaaa aagcttactg gcttaactat gcggcatcag agcagattgt actgagagtg
4921 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
4981 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
5041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
5101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
5161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
5221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
5281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
5341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
5401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
5461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
5521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
5581 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
5641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
5701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
5761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
5821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
5881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
5941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
6001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
6061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
6121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
6181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
6241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
6301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
6361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
6421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
6481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
6541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
6601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
6661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
6721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
6781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
6841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
6901 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
6961 cgtatcacga ggccctttcg tcttcaagaa ttagcttggc cattgcatac gttgtatcca
7021 tatcataata tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga
7081 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
7141 gagt
//
