| 质粒名称: | pPKm-145 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 90505 |
| 存储实验室: | Todd Coleman |
|
载体骨架基本信息 |
|
| 载体名称: | pSIN |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pPKm-145 was a gift from Todd Coleman (Addgene plasmid # 90505 ; http://n2t.net/addgene:90505 ; RRID:Addgene_90505) |
| 参考文献部分: | Biosynthesis of Orthogonal Molecules Using Ferredoxin and Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded PhyB Optogenetics. Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP. ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 10.1021/acssynbio.7b00413. Epub 2018 Jan 24. 10.1021/acssynbio.7b00413 PubMed 29301067 |
买家导航
pPKm-145载体 pPKm145质粒基本信息
pPKm-145质粒图谱 pPKm145载体图谱和pPKm-145载体序列 pPKm145质粒序列多克隆位点信息
pPKm-145质粒 pPKm145载体简介
pPKm-145质粒序列 pPKm145载体序列
LOCUS pPKm-145 7589 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Empty plasmid, pSIN-EF1-alpha-IRES-puro.
ACCESSION .
VERSION .
KEYWORDS pPKm-145
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7589)
AUTHORS Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS,
Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
TITLE Biosynthesis of Orthogonal Molecules Using Ferredoxin and
Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded
PhyB Optogenetics.
JOURNAL ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi:
10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
PUBMED 29301067
REFERENCE 2 (bases 1 to 7589)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7589
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..1180
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 233..1171
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1128..1148
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 1297..1870
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1464..1481)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1691..1710
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 1903..2502
/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(1903..1922)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2399..2419
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 2705..2938
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(3089..3106)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3095..3319
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
regulatory 3362..3371
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter 3415..3519
/gene="bla"
/label=AmpR promoter
CDS 3520..4380
/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 complement(3738..3757)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4551..5139
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5040..5059
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
LTR 5298..5931
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 5978..6103
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6596..6829
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 7345..7462
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
61 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
121 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
181 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
241 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
301 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
361 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
421 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
481 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
541 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
601 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
661 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
721 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
781 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
841 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
901 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
961 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
1021 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
1081 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
1141 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattgatc cttcgaacta
1201 gaattcgaag cttgagctcg agatctgagt ccggtagcgc tagttcgaag gatccgcatg
1261 catctagggc ggccaattcc gcccctctcc cccccccccc tctccctccc ccccccctaa
1321 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
1381 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
1441 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
1501 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
1561 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
1621 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
1681 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
1741 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
1801 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
1861 acgatgataa gcttgccaca acccacaagg agacgacctt ccatgaccga gtacaagccc
1921 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
1981 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
2041 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
2101 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
2161 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
2221 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
2281 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
2341 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
2401 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
2461 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gataataggc ggccgctcga
2521 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
2581 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
2641 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
2701 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
2761 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
2821 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
2881 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
2941 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
3001 agtgagaggc cttgacatta taatagattt agcaggaatt gaactaggag tggagcacac
3061 aggcaaagtt ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
3121 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
3181 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
3241 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
3301 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctgggggcg cgcccctcga
3361 ggccgccatg gtcatagctg tttgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
3421 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcac gagacaataa
3481 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
3541 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
3601 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
3661 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
3721 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
3781 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
3841 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
3901 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
3961 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
4021 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
4081 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
4141 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
4201 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
4261 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
4321 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
4381 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
4441 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
4501 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
4561 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
4621 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
4681 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
4741 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
4801 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
4861 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
4921 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
4981 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
5041 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
5101 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
5161 ttacggttct ggccttttgc acggaccgcc actgccaatt acctgtggtt tcatttactc
5221 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
5281 actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga
5341 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
5401 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
5461 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
5521 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
5581 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
5641 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
5701 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
5761 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
5821 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
5881 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
5941 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
6001 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
6061 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
6121 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
6181 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
6241 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
6301 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
6361 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
6421 taagaccacc gcacagcaac cggccgctga tcttcagacc tggaggagga gatatgaggg
6481 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
6541 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
6601 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
6661 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
6721 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
6781 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
6841 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
6901 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
6961 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
7021 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
7081 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
7141 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
7201 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
7261 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc
7321 acaaatggca gtattcatcc acaattttaa aagaaagggg gggattgggg ggtacagtgc
7381 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
7441 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggat
7501 cgataagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
7561 accttctagg tcttgaaagg agtgggaat
//
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