| 质粒名称: | pPGK-Cre-bpA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11543 |
| 存储实验室: | Klaus Rajewsky |
|
载体骨架基本信息 |
|
| 载体名称: | XhopKS |
| 载体出品公司: | Stratagene |
| 空载体大小: | 2906 bp |
| 完整质粒大小: | 4870 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，Cre/Lox |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 磷酸甘油酸激酶1启动子 |
| 物种来源: | 噬菌体P1 |
| 插入大小: | 1944 bp |
| 基因突变: | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Sall(未破坏) |
| 3’克隆位点: | Not I(未破坏) |
| 5’测序引物 | n/a |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pPGK-Cre-bpA was a gift from Klaus Rajewsky (Addgene plasmid # 11543 ; http://n2t.net/addgene:11543 ; RRID:Addgene_11543) |
| 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 pPGK-Cre-bpA 4870 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pPGK-Cre-bpA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4870)
TITLE Rajewsky pPGK-Cre-bpA
REFERENCE 2 (bases 1 to 4870)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4870
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 52..156
/gene="bla"
/label=AmpR promoter
CDS 157..1017
/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(375..394)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1188..1776
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1677..1696
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1930..1947
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2064..2085
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2100..2130
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2138..2154
/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 2143..2165
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2162..2178
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2162..2178
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2197..2217
/label=T3
/note="T3 promoter, forward primer"
promoter 2199..2217
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 2246..2262
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 2247..2263
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2323..2346)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 2712..2731
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 2804..2824
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2825..3853
/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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
GD"
primer_bind complement(3877..3893)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3877..3893)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(3907..3924)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3913..4137
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(4228..4247)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4229..4247)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4257..4274)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4257..4273)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4266..4288)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4415..4870
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4502..4521)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4712..4733
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 atgcaatcct actatgcaat gtatccaggt ggcacttttc ggggaaatgt gcgcggaacc
61 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
1021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
1081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
1141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
1201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
1261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
1321 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
1381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
1441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
1501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
1561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
1621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
1681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
1741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
1801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
1861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
1921 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
1981 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
2041 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
2101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
2161 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
2221 caaaagctgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgatatcgaa
2281 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
2341 ccgctggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
2401 gtagcgccaa ccggctccgt tctttggtgg ccccttcgcg ccacttctac tcctccccta
2461 gtcaggaagt ttcccccagc aagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
2521 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
2581 ggggcagcgg ccaatagcag ctttgttcct tcgctttctg ggctcagagg ctgggaaggg
2641 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg gcgggcgccc gaaggtcctc
2701 ccgaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
2761 ctcatctccg ggcctttcga cctgcagctc gaggtcgacc atgcccaaga agaagaggaa
2821 ggtgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
2881 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
2941 tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
3001 ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc
3061 gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg
3121 tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat
3181 ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac
3241 tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa
3301 tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat
3361 cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac
3421 gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact
3481 ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg
3541 ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc
3601 cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg
3661 tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc
3721 ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat
3781 tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga
3841 agatggcgat tagccattaa cgcgtaaatg attgcagatc cactagttct agagctcgct
3901 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
3961 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
4021 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
4081 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
4141 ctgaggaaag aaccagctgg ggctcgagat ccactagttc tagcctcgag gctagagcgg
4201 ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattacgc gcgctcactg
4261 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
4321 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
4381 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
4441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
4501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
4561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
4621 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
4681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
4741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
4861 cttacaattt
//
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