| 质粒名称: | ORFtag_TetR-SD_Frame2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 220982 |
| 存储实验室: | Alexander Stark |
|
载体骨架基本信息 |
|
| 载体名称: | ORFtag backbone |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 7682bp |
| 载体类型: | 哺乳细胞表达, Retroviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | TetR repressor / TetR-splice donor in Frame2 |
|
文献引用方法 |
|
| 材料和方法部分: | ORFtag_TetR-SD_Frame2 was a gift from Alexander Stark (Addgene plasmid # 220982 ; http://n2t.net/addgene:220982 ; RRID:Addgene_220982) |
买家导航
ORFtag_TetR-SD_Frame2载体质粒基本信息
ORFtag_TetR-SD_Frame2质粒图谱载体图谱和ORFtag_TetR-SD_Frame2载体序列质粒序列多克隆位点信息
ORFtag_TetR-SD_Frame2质粒载体简介
ORFtag_TetR-SD_Frame2质粒序列载体序列
LOCUS ORFtag_TetR-SD_Frame2 7682 bp DNA circular SYN 14-APR-2025
DEFINITION ORFtag viral retroviral vector containing a constitutively active
promoter and a TetR tag followed by a splice donor sequence in
Frame2.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7682)
TITLE Proteome-scale tagging and functional screening in mammalian cells
by ORFtag
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7682)
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"Unpublished"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: ORFtag_TetR-SD_Frame2
FEATURES Location/Qualifiers
source 1..7682
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 143..162
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 396..413
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(494..513)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter complement(535..864)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 549..684
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 849..869
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind 1005..1027
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter 1481..1980
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(1516..1539)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 1913..1932
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 1993..2787
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2047..2066)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2657..2676
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
misc_feature join(2793..3366,3367..3369,3370..3378)
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2959..2976)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3186..3205
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3394..3414
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4072..4107
/codon_start=1
/product="nanobody epitope tag corresponding to amino acids
16-27 of human beta-catenin (Traenkle et al., 2015)"
/label=BC2 tag
/translation="PDRKAAVSHWQQ"
CDS 4111..4176
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS complement(4376..4792)
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind complement(4392..4408)
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(4424..4446)
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
misc_feature 4993..5192
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
LTR 5255..5430
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
promoter complement(5431..5634)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(5460..5480)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 5635..6014
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(6114..6132)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6200..6304
/gene="bla"
/label=AmpR promoter
CDS join(6305..6373,6374..7165)
/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 complement(6523..6542)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin join(7336..7682,1..242)
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
61 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
301 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
361 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
421 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
481 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtaa gctttttgca
541 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
601 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
661 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
721 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
781 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
841 ctttccacac cctaactgac acacattcca cagccggatc tacactccgc tatcgctacg
901 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
961 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
1021 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ccagcttacc tcccggtggt
1081 gggtcggtgg tccctgggca ggggtctccc gatcccggac gagcccccaa atgaaagacc
1141 cccgctgacg ggtagtcaat cactcagagg agaccctccc aaggaacagc gagaccacaa
1201 gtcggatgca actgcaagag ggtttattgg atacacgggt acccgggcga ctcagtcaat
1261 cggaggactg gcgccccgag tgaggggttg tgctagcttg ccaaacctac aggtggggtc
1321 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatcg atggtcaagt
1381 tccttctggt tctggctctg ctcatcgtcc tgcaggaccg gacgtgactg gagttcagac
1441 gtgtgctctt ccgatctcat gttaactcga gaattctacc gggtagggga ggcgcttttc
1501 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
1561 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
1621 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
1681 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
1741 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
1801 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
1861 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
1921 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
1981 gaattcgcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
2041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
2101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
2161 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
2221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
2281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
2341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
2401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
2461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
2521 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
2581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
2641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
2701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
2761 tatcgccttc ttgacgagtt cttctgacgc gtcccctctc cctccccccc cctaacgtta
2821 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
2881 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
2941 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
3001 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
3061 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
3121 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
3181 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
3241 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
3301 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
3361 gataatatcg ccacaaccag atctgccacc atgcccaaaa agaagaggaa agtgatgtct
3421 agactggaca agagcaaagt cataaactct gctctggaat tactcaatga agtcggtatc
3481 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
3541 tggcacgtga agaacaagcg ggccctgctc gatgccctgg caatcgagat gctggacagg
3601 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
3661 gccaagtcat tccgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
3721 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
3781 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
3841 ggctgcgtat tggaggatca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
3901 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgacca tcagggagcc
3961 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
4021 tgcgaaagcg gcgggccgga tatcggaggc ggatcaggtg gcggtagcgt ccccgatcgt
4081 aaggctgcgg tctctcattg gcaacaggac gactataagg accacgacgg agactacaag
4141 gatcatgata ttgattacaa agacgatgac gataaggcag gtaagtaatg cttttttttt
4201 tctcgctctc atttttcaaa aacccacata aaaattgagg aaagggaaga attgttttct
4261 ccttccagca cctcgtaatt tgacccgact gatggttccc attagtcaca taaagctgta
4321 gtcaagtaca gacgtcccgc ggcttcgata ccgtcacgaa gcaacggatc cggccggcgc
4381 ctagagaagg agtgagggct ggataaaggg aggatcgagg cggggtcgaa cgaggaggtt
4441 caagggggag agacggggcg gatggaggaa gaggaggcgg aggcttaggg tgtacaaagg
4501 gcttgaccca gggagggggg tcaaaagcca aggcttccca ggtcacgatg taggggacct
4561 ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat cttaacctgg gtgatgaggt
4621 ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa ggttggccat tctgcagagc
4681 agaaggtaac ccaacgtctc ttcttgacat ctaccgactg gttgtgagcg atccgctcga
4741 catctttcca gtgacctaag gtcaaactta agggagtggt aacagtctgg ccctaatttt
4801 cagacaaata cagaaacaca gtcagacaga gacaacacag aacgatgctg cagcgctgca
4861 gcagacaaga cgcgcggctt cggttccaaa ccgaaagcaa aaattcagac ggaggcggga
4921 actgttttag gttctcgtct cctaccagaa ccacatatcc tgacggggtc ggattccaca
4981 tcgactccct tcctcaggtc gggccacaaa aacggccccc aaagtccctg ggacgtctcc
5041 cagggttgcg gccgggtgtt cagaactcgt cagttccacc acgggtccgc cagatacaga
5101 gctagttagc taactagtac agacgcaggc gcataacatc aaacatagac actagacaat
5161 cggacagaca cagataagtt gctggccagc ttacctcccg gtggtgggtc ggtggtccct
5221 gggcaggggt ctccaaatcc cggacgagcc cccaaatgaa agacccccgt cgtgggtagt
5281 caatcactca gaggagaccc tcccaaggaa cagcgagacc acgattcgga tgcaaacagc
5341 aagaggcttt attgggaata cgggtacccg ggcgacgcag tctatcggaa gactggcgcg
5401 ccgagtgagg ggttgtgggc tcttttattg agctctgctt atatagacct cccaccgtac
5461 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
5521 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
5581 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
5641 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
5701 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
5761 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
5821 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
5881 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
5941 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt
6001 caataatcaa tgtcaacatg gcggtaatgt tggacatgag ccaatataaa tgtacatatt
6061 atgatatgga tacaacgtat gcaatggcca agctaattct tgaagacgaa agggcctcgt
6121 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
6181 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
6241 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
6301 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
6361 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
6421 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
6481 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
6541 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
6601 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
6661 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
6721 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
6781 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
6841 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
6901 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
6961 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
7021 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
7081 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
7141 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
7201 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
7261 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
7321 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
7381 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
7441 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
7501 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
7561 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
7621 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
7681 ct
//
