| 质粒名称: | pMXspuro-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74203 |
| 存储实验室: | Jayanta Debnath |
|
载体骨架基本信息 |
|
| 载体名称: | pMXs-puro |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6900 bp |
| 完整质粒大小: | 7600 bp |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | 嘌呤霉素 |
| 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: | GFP |
| 插入大小: | 717 bp |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(销毁时未知) |
| 3' 克隆位点: | XhoI (销毁时未知) |
| 5' 测序引物 | GACGGCATCGCAGCTTGGATACAC |
|
文献引用方法 |
|
| 材料和方法部分: | pMXspuro-GFP was a gift from Jayanta Debnath (Addgene plasmid # 74203 ; http://n2t.net/addgene:74203 ; RRID:Addgene_74203) |
| 参考文献部分: | NBR1 enables autophagy-dependent focal adhesion turnover. Kenific CM, Stehbens SJ, Goldsmith J, Leidal AM, Faure N, Ye J, Wittmann T, Debnath J. J Cell Biol. 2016 Feb 29;212(5):577-590. Epub 2016 Feb 22. 10.1083/jcb.201503075 PubMed 26903539 |
买家导航
pMXspuroGFP载体基本信息
pMXspuroGFP载体图谱质粒图谱和多克隆位点信息
pMXspuroGFP载体简介
pMXspuroGFP载体序列
LOCUS pMXspuro-GFP 6344 bp ds-DNA circular SYN 09-DEC-2019
DEFINITION Retroviral control vector containing GFP insert.
ACCESSION .
VERSION .
KEYWORDS pMXspuro-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6344)
AUTHORS Kenific CM, Stehbens SJ, Goldsmith J, Leidal AM, Faure N, Ye J,
Wittmann T, Debnath J
TITLE NBR1 enables autophagy-dependent focal adhesion turnover.
JOURNAL J Cell Biol. 2016 Feb 29;212(5):577-590. Epub 2016 Feb 22.
PUBMED 26903539
REFERENCE 2 (bases 1 to 6344)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 9, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6344
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..375
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind 313..329
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
CDS 407..1126
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(452..473)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(713..732)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1060..1081
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1160..1180)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1165..1494
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1345..1480
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1407..1426
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 1504..2103
/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(1504..1523)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2000..2020
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 2330..2799
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(2929..2946)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3100..3688)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3180..3199)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3852..4712)
/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 4475..4494
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4713..4817)
/gene="bla"
/label=AmpR promoter
LTR 4847..5440
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 5503..5860
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 5919..6335
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 6265..6287
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 6303..6319
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
ORIGIN
1 ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga ccctgacatg
61 acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta cttagtccag
121 cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg accggtggta
181 cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac taagaaccta
241 gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc cctcaaagta
301 gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga
361 ccatcctcta gactgccgga tctagctagt taattaagga tctaccatgg tgagcaaggg
421 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
481 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
541 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
601 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
661 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
721 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
781 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
841 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
901 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
961 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
1021 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
1081 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtgactcg agcggccgcc
1141 agcacagtgg tcgacctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
1201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
1261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
1321 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
1381 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
1441 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
1501 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
1561 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
1621 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
1681 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
1741 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
1801 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
1861 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
1921 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
1981 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
2041 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
2101 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
2161 tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
2221 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
2281 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
2341 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
2401 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
2461 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
2521 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
2581 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
2641 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
2701 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
2761 cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt
2821 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
2881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
2941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
3001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
3061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
3121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
3181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
3241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
3301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
3361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
3421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
3481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
3541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
3601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
3661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
3721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
3781 cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact
3841 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
3901 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
3961 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
4021 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
4081 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
4141 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
4201 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
4261 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
4321 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
4381 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
4441 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
4501 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
4561 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
4621 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
4681 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
4741 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
4801 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcataatg aaagacccca
4861 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
4921 aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
4981 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
5041 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
5101 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
5161 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
5221 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
5281 aacccctcac tcggcgcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
5341 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
5401 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
5461 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
5521 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
5581 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tcggaacacc
5641 cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg gcccgacctg
5701 agtccaaaaa tcccgatcgt tttggactct ttggtgcacc cccctaatag gagggatatg
5761 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
5821 cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc
5881 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
5941 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
6001 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
6061 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
6121 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
6181 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
6241 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
6301 ccctttatcc agccctcact ccttctctag gcgcccccat atgg
//
