| 质粒名称: | pCDH-EF1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 72266 |
| 存储实验室: | Kazuhiro Oka |
|
载体骨架基本信息 |
|
| 载体名称: | pCDH-CMV-MCS |
| 载体出品公司: | Systembio |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7133 bp |
| 完整质粒大小: | -- |
| 载体修饰: | ClaI/SalI fragment was replaced with EF1 + new MCS |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 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: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | GCACTTGATGTAATTCTCC |
| 3' 测序引物 | CAACACCACGGAATTGTCAG |
|
文献引用方法 |
|
| 材料和方法部分: | pCDH-EF1 was a gift from Kazuhiro Oka (Addgene plasmid # 72266 ; http://n2t.net/addgene:72266 ; RRID:Addgene_72266) |
买家导航
pCDHEF1载体基本信息
pCDHEF1载体图谱质粒图谱和多克隆位点信息
pCDHEF1载体简介
pCDHEF1载体序列
LOCUS pCDH-EF1 7139 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Express gene of interest under EF-1 promoter.
ACCESSION .
VERSION .
KEYWORDS pCDH-EF1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7139)
TITLE lenti vector
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7139)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7139
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 234..414
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 458..583
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1076..1309
/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 1804..1921
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2019..3197
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2250..3188
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3145..3165
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 3234..3822
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3287..3307)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3705..3716)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3896..4129
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4201..4322
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4238..4257)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4292..4311
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4341..4476
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4403..4422
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4514..4530)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4514..4530)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4527..4549)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4538..4554
/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)."
promoter complement(4562..4592)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4607..4628
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4745..4762)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4916..5504)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4996..5015)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5675..6535)
/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 6298..6317
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6536..6640)
/gene="bla"
/label=AmpR promoter
primer_bind 6708..6726
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6764..6786)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6886..6905
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7099..7121
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7113..7130
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7114..7130
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
1921 tatcgatatc gatagctttg caaagatgga taaagtttta aacagagagg aatctttgca
1981 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
2041 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
2101 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
2161 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
2221 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
2281 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
2341 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
2401 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
2461 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
2521 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
2581 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
2641 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
2701 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
2761 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
2821 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
2881 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
2941 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
3001 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
3061 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
3121 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
3181 catttcaggt gtcgtgagga attggatccc cgggcggccg cgaattcgtc gacaatcaac
3241 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
3301 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
3361 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
3421 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
3481 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
3541 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
3601 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
3661 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
3721 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
3781 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca
3841 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
3901 gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg
3961 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
4021 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
4081 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
4141 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
4201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
4261 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
4321 tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc
4381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
4441 tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa
4501 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
4561 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
4621 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
4741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
4801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
4981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
5041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
5101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
5161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
5221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
5281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
5341 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
5401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
5461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
5521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
5581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
5641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
5701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
5761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
5821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
5881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
5941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
6001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
6061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
6121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
6181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
6241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
6301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
6361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
6421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
6481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
6541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
6601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
6661 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
6721 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
6781 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
6841 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
6901 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
6961 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
7021 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
7081 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
