| 质粒名称: | pBS513 EF1alpha-cre |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11918 |
| 存储实验室: | Brian Sauer |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6600 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，Cre/Lox |
| 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: | cre |
| 物种来源: | bacteriophage P1 |
| 插入大小: | 1100 bp |
| 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 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pBS513 EF1alpha-cre was a gift from Brian Sauer (Addgene plasmid # 11918 ; http://n2t.net/addgene:11918 ; RRID:Addgene_11918) |
| 参考文献部分: | GFPcre fusion vectors with enhanced expression. Le Y, Miller JL, Sauer B. Anal Biochem. 1999 Jun 1. 270(2):334-6. 10.1006/abio.1999.4110 PubMed 10334853 |
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 pBS513 EF1alpha-cre 7927 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBS513 EF1alpha-cre
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7927)
AUTHORS Le Y, Miller JL, Sauer B
TITLE GFPcre fusion vectors with enhanced expression.
JOURNAL Anal Biochem. 1999 Jun 1. 270(2):334-6.
PUBMED 10334853
REFERENCE 2 (bases 1 to 7927)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7927
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..1032
/codon_start=1
/product="site-specific recombinase"
/label=Cre
/note="Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP sequences."
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
RBS 2684..2692
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
primer_bind complement(3486..3503)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3486..3502)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3495..3517)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3715..4170
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3802..3821)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4012..4033
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4187..4206)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4306..4328
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4366..4384)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4452..4556
/gene="bla"
/label=AmpR promoter
CDS 4557..5417
/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(4775..4794)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5588..6176
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6077..6096
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6330..6347
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6464..6485
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6500..6530
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6538..6554
/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 6543..6565
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6562..6578
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6562..6578
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 6677..7855
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 6908..7846
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 7803..7823
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
ORIGIN
1 atgtccaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt
61 gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat
121 acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac
181 cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg
241 cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt
301 cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc
361 cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact
421 gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat
481 ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc
541 agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg
601 aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg
661 gtcgagcgat ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc
721 cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc
781 ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt
841 cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc
901 cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt
961 gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa
1021 gatggcgatt agccattaac gcgtaaatga ttgctataat tatttgatat ttatggtgac
1081 atatgagaaa ggatttcaac atcgacggaa aatatgtagt gctgtctgta agcactaata
1141 ttcagtcgcc agccgtcatt gtcactgtaa agctgagcga tagaatgcct gatattgact
1201 caatatccgt tgcgtttcct gtcaaaagta tgcgtagtgc tgaacatttc gtgatgaatg
1261 ccaccgagga agaagcacgg cgcggttttg ctaaagtgat gtctgagttt ggcgaactct
1321 tgggtaaggt tggaattgtc gagggatctc ggaatggacc ccaactgctc ctgctccacc
1381 ggtaagactc ccgatccttg gtctttagaa taccaagttg ggaccgcaga gcggaatccc
1441 cgagttgtag aggcttggcg ggaataggca cctttagttg gcgattcatt ccggttcttt
1501 ctagaatccg ctcttgcaaa agccttcatt agttacgagt attgtcgaac gggtcctttg
1561 gcggggttgg ggctaggatt tagacgcgca aatgtccggt tcctgatcac ccagttagtg
1621 gggacatctg ggttgagtcc caggcattac taaacttact gtgaattgct tgaattaaga
1681 aagaggtgaa ggacctttat gtcttgggac tcaaagacat aatccctgac ttaacctgtg
1741 aggagaaaag tggggctagg ctccctgcag ctccgaggag gacttagtga actgagccgg
1801 gactcgtggt gttggccact gctgtaatgc tgcctccctc atgctgtctt ctttctcctc
1861 cccaggcggc tcctgcactt gcaccagctc ctgcgcctgc aagaactgca agtgcacctc
1921 ctgcaagaag agtgagttgg gacaccttgg gtggcggcta aggctagggg cggggaactc
1981 ctacaaaact ggctctgaga aatgtccttt gcttcccgga ggccattgta ttgtctcggg
2041 gacagaacta tacagagaac tatttaaaaa aaccgaggtc ttctctgttg gggacaggaa
2101 gcagaggtct tcagccaggc tgacctcttc ctcctccttt ctaggctgct gctcctgctg
2161 tcccgtgggc tgctccaaat gtgcccaggg ctgtgtctgc aaaggcgccg cggacaagtg
2221 cacgtgctgt gcctgatgtg acgaacagcg ctgccaccac gtgtaaatag tatcggacca
2281 acccagcgtc ttcctataca gttccaccct gtttactaaa cccccgtttt ctaccgagta
2341 cgtgaataat aaaagcctgt ttgagtctaa ctctggtttt cttggtgtgg tttggcaata
2401 agaaactggg gtgacttgat agtctgggga tctggttttg gaccccctcg tgcctttacc
2461 tcccgcctct ggccctcaca gaggggtaat gtctttgggt aaagccaagc tatcccataa
2521 gcttcctcat ggaaaacagc tgtgtgcctg gtcaggtctt gtgttagggg aagggaggga
2581 gggagggtgg tcctggaaga cttgatgtcc catcctggaa gcacagaaaa tcttatcatt
2641 tatattctct ttggattctg gtttcctctc acattgttgg gtataaggag gtgggtatga
2701 aaattaacat agggtgtctc gtatgctagg cagggactaa cactgaacta gattccagag
2761 gggctttctt ggctgttttg acacaagttg cttaccttgt cctagaactc attctgcagt
2821 ccagtctgac cttgagtttt tgattctcct gcctcagctt cccaagtacc tgggattaca
2881 ggagtgactg tgtcaccagg ccatctccta ttcagccacc aaatgcagac actattgcat
2941 atgccatcaa gattttgctg aaaggaccct gataaagctg tctcctgtga ggctttgcca
3001 gtgcccagca aacacagaag tggatgctca cagtcagcta ttggatggaa cacagggccc
3061 ccaatggagg agctagagaa agtacccaag gagctaaagg gatctgcaac cctataggtg
3121 gaacagcaat ctgaactaac cagtaccccc tgaactcgtg tatctagctt cgtatgtagc
3181 agaagatggc ctagttggcc atcactggga agagaggacc cttggtctag caaactatat
3241 gccccagttc aggggaaagc cagggccaag aagtgggagt gagtgggtag gggagcgggg
3301 tgggggtagg ggtggggtat aggggacttt tgggatagca tttgaaatat aaacgaagaa
3361 aatatccaaa aaaaaaaaga tggaaaaaaa agtgataaag agcgtgggtc ctgcctgtaa
3421 tcaagatatt tgagatgtag cagcaggaag gccaggaatt ggtcgagggt accgagctcg
3481 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
3541 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
3601 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
3661 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
3721 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
3781 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
3841 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
3901 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
3961 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
4021 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
4081 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
4141 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
4201 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
4261 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
4321 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
4381 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
4441 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
4501 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
4561 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
4621 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
4681 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
4741 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
4801 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
4861 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
4921 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
4981 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
5041 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
5101 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
5161 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
5221 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
5281 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
5341 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
5401 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
5461 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
5521 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
5581 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
5641 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
5701 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
5761 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
5821 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
5881 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
5941 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
6001 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
6061 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
6121 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
6181 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
6241 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
6301 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
6361 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
6421 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
6481 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
6541 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctttgca
6601 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
6661 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
6721 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
6781 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
6841 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
6901 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
6961 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
7021 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
7081 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
7141 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
7201 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
7261 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
7321 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
7381 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
7441 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
7501 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
7561 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
7621 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
7681 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
7741 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
7801 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat
7861 tggatccaga ttcgaggggc agagccgatc ctgtacactt tacttaaaac cattatctga
7921 gtgtgaa
//
