| 质粒名称: | pQCXIP-EGFP-F |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 73014 |
| 存储实验室: | Michael Grusch |
|
载体骨架基本信息 |
|
| 载体名称: | pQCXIP |
| 载体出品公司: | Clontech |
| 空载体大小: | 7200bp |
| 完整质粒大小: | 8050bp |
| 载体类型: | 哺乳细胞表达, Retroviral |
| 筛选标记: | Puromycin |
| 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 |
|
| 插入基因名称: | EGFP-F、farnesylated enhanced green fluorescent protein |
| 物种来源: | Aequorea victoria |
| 插入大小: | 850bp |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | LNCX |
| 3’测序引物 | IRES rev |
|
文献引用方法 |
|
| 材料和方法部分: |
pQCXIP-EGFP-F was a gift from Michael Grusch (Addgene plasmid # 73014 ; http://n2t.net/addgene:73014 ; RRID:Addgene_73014)
|
买家导航
pQCXIP-EGFP-F载体质粒基本信息
pQCXIP-EGFP-F质粒图谱载体图谱和pQCXIP-EGFP-F载体序列质粒序列多克隆位点信息
pQCXIP-EGFP-F质粒载体简介
pQCXIP-EGFP-F质粒序列载体序列
LOCUS pQCXIP-EGFP-F 8027 bp DNA circular SYN 14-APR-2025
DEFINITION Retroviral expression of EGFP-F.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8027)
TITLE Grusch plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8027)
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:
"Unpublished"
COMMENT Sequence Label: pQCXIP-EGFP-F
FEATURES Location/Qualifiers
source 1..8027
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 309..512
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 513..688
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 751..950
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1151..1567
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1497..1519
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1535..1551
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
enhancer 1604..1907
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1908..2111
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2108..2132
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 2323..2332
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(2329..2331,2332..2334,2335..3045)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(2329..2331,2332..2334,2335..3045)
/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(2374..2395)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2635..2654)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2982..3003
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature join(3154..3728,3729..3731,3732..3740)
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3321..3338)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3548..3567
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3760..4359
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3760..3779)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4256..4276
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 4733..5158
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
primer_bind complement(5261..5283)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5419..5439)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5424..5753
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5604..5739
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5775..5794
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(5875..5892)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6046..6634)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6126..6145)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6805..7596,7597..7665))
/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 7428..7447
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7666..7770)
/gene="bla"
/label=AmpR promoter
primer_bind 7838..7856
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
enhancer join(7956..8027,1..308)
/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 gcccgcctgg cattatgccc
241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
1561 aggcgccgga attgaagatc tgggggatcg atcctctaga gtccgttaca taacttacgg
1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
1921 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
2101 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
2161 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
2221 cggacctgca ggaattgatc cgcggccgca ccggtaggcc tcgtacgctt aattaacgga
2281 tccggtaccg aattcgcggc cgcactcgag atgctaccgg tcgccaccat ggtgagcaag
2341 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
2401 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
2461 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
2521 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
2581 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
2641 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
2701 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
2761 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
2821 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
2881 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
2941 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
3001 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
3061 aagctgaacc ctcctgatga gagtggcccc ggctgcatga gctgcaagtg tgtgctctcc
3121 tgaggatccg gaattccgcc cccccccccc ccccccctct ccctcccccc cccctaacgt
3181 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
3241 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
3301 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
3361 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
3421 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
3481 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
3541 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
3601 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
3661 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
3721 atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta caagcccacg
3781 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
3841 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
3901 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
3961 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
4021 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
4081 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
4141 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
4201 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
4261 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
4321 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
4381 cgcccgaccg aaaggagcgc acgaccccat ggctccgacc gaagccaccc ggggcggccc
4441 cgccgacccc gcacccgccc ccgaggccca ccgactctag tcgagggctg cagcgctgca
4501 gaggccgagt gcagaactgc tccaaaggga cctcaaggct ttccgaggga cactaggctg
4561 actccatcga gccagtgtag agataagctt atcgattagt ccaatttgtt aaagacagga
4621 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt
4681 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
4741 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
4801 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagggtcga
4861 ccctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
4921 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
4981 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
5041 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
5101 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
5161 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt
5221 aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
5281 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
5341 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
5401 gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
5461 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
5521 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
5581 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
5641 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
5701 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta
5761 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
5821 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
5881 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
5941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
6001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
6061 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
6121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
6181 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
6241 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
6301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
6361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
6421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
6481 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
6541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
6601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
6661 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
6721 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
6781 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
6841 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
6901 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
6961 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
7021 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
7081 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
7141 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
7201 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
7261 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
7321 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
7381 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
7441 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
7501 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
7561 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
7621 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
7681 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
7741 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
7801 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
7861 tcgtcttcaa gaattagctt ggccattgca tacgttgtat ccatatcata atatgtacat
7921 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
7981 atagtaatca attacggggt cattagttca tagcccatat atggagt
//
