| 质粒名称: | pUltra-hot |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24130 |
| 存储实验室: | pRRL |
|
载体骨架基本信息 |
|
| 载体名称: | Malcolm Moore |
| 载体出品公司: | Didier Trono |
| 空载体大小: | 7000 bp |
| 完整质粒大小: | 8317 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 启动子: | -- |
| 标签/融合蛋白: | mCherry（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XbaI/NheI(未破坏) |
| 3’克隆位点: | BamHI/BcII (未破坏) |
| 5’测序引物 | hUBCpro-F |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pUltra-hot was a gift from Malcolm Moore (Addgene plasmid # 24130 ; http://n2t.net/addgene:24130 ; RRID:Addgene_24130) |
| 参考文献部分: | -- |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pUltra-hot 8317 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION 3rd generation Lentiviral vector for bi-cistronic expression of
mCherry and the gene of interest.
ACCESSION .
VERSION .
KEYWORDS pUltra-hot
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8317)
TITLE pUltra series
REFERENCE 2 (bases 1 to 8317)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8317
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1..589
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 490..509
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 743..760
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 877..898
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 913..943
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 951..967
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 956..978
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 975..991
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 975..991
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 1010..1030
/label=T3
/note="T3 promoter, forward primer"
promoter 1012..1030
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1058..1284
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 1285..1465
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1512..1637
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2130..2363
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2548..2592
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2890..3007
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 3072..4283
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 4120..4139
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
regulatory 4316..4325
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4322..5029
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(4467..4485)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 4735..4754
/label=mCherry-F
/note="mCherry, forward primer"
CDS 5039..5095
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 5117..5170
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
misc_feature 5214..5802
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5267..5287)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5685..5696)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5893..6126
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6198..6319
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6235..6254)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6289..6308
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6359..6494
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6421..6440
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(6514..6533)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6515..6533)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6543..6560)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6543..6559)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6552..6574)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 6701..7156
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6788..6807)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6998..7019
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7182..7286
/gene="bla"
/label=AmpR promoter
CDS 7287..8147
/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(7505..7524)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctcact
541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
2821 attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt
2881 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
2941 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
3001 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta acccgtgtcg
3061 gctccagatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac
3121 ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt ccgcccggac
3181 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
3241 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
3301 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
3361 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
3421 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
3481 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
3541 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
3601 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
3661 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
3721 cttgaggcct tcgctaatgc gggaaagctc ttattcgggt gagatgggct ggggcaccat
3781 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
3841 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
3901 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
3961 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
4021 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
4081 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
4141 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
4201 catttgggtc aatatgtaat tttcagtgtt agactagtaa attgtccgct aaattctggc
4261 cgtttttggc ttttttgtta gacgaagctt gggctgcagg tccgatccac cggtcgccac
4321 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
4381 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
4441 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
4501 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
4561 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
4621 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
4681 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
4741 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
4801 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
4861 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
4921 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
4981 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg gatctggagc
5041 aacaaacttc tcactactca aacaagcagg tgacgtggag gagaatcccg ggccttctag
5101 aggatccggc tccggcgagg gcaggggaag tctactaaca tgcggggacg tggaggaaaa
5161 tcccggccca gctagcgtgt gatcagaatt cgttccggag tcgtcgactc gacaatcaac
5221 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
5281 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
5341 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
5401 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
5461 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
5521 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
5581 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
5641 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
5701 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcat cttcgccttc
5761 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattcgagct
5821 cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
5881 aaaagggggg actggaaggg ctacgtaact cccaacgaag acaagatctg ctttttgctt
5941 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
6001 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
6061 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
6121 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
6181 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
6241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
6301 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
6361 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
6421 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
6481 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
6541 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
6601 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
6661 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
6721 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
6781 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
6841 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
6901 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
6961 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
7021 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
7081 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
7141 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
7201 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
7261 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
7321 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
7381 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
7441 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
7501 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
7561 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
7621 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
7681 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
7741 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
7801 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
7861 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
7921 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
7981 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
8041 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
8101 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
8161 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
8221 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
8281 gagcgtcaga ccccgtagaa aagatcaaag gatcttc
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