| 质粒名称: | p11-LacY-wtx1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 69056 |
| 存储实验室: | Huimin Zhao |
|
载体骨架基本信息 |
|
| 载体名称: | pBAD18 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4600 bp |
| 完整质粒大小: | 4600 bp |
| 筛选标记: | 氨苄青霉素 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | ccdB |
| 启动子 | pBAD |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI（销毁后未知） |
| 3' 克隆位点: | SphI（销毁后未知） |
| 5' 测序引物 | catgca gctagc GGAGTG aaacgatgcagttt aaggtttacacctataaaaga |
| 3' 测序引物 | agtacg gcatgc cgtatg tctaga ttatattccccaaaacatcaggttaat |
|
文献引用方法 |
|
| 材料和方法部分: | p11-LacY-wtx1 was a gift from Huimin Zhao (Addgene plasmid # 69056 ; http://n2t.net/addgene:69056 ; RRID:Addgene_69056) |
| 参考文献部分: | A highly sensitive selection method for directed evolution of homing endonucleases. Chen Z, Zhao H. Nucleic Acids Res. 2005 Oct 6;33(18):e154. 10.1093/nar/gni148 PubMed 16214805 |
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 p11-LacY-wtx1 6092 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION The reporter plasmid p11-LacY-wtx1 encodes the toxin CcdB gene under
the inducible BAD promoter and a single copy of the endonuclease
cleavage site which is readily extendable to multiple copies..
ACCESSION .
VERSION .
KEYWORDS p11-LacY-wtx1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6092)
AUTHORS Chen Z, Zhao H
TITLE A highly sensitive selection method for directed evolution of homing
endonucleases.
JOURNAL Nucleic Acids Res. 2005 Oct 6;33(18):e154.
PUBMED 16214805
REFERENCE 2 (bases 1 to 6092)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6092
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(95..973)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 1000..1284
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 1207..1226
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 1316..1621
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 1500..1519
/label=ccdB-fwd
/note="ccdB gene, forward primer"
primer_bind complement(1716..1733)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 1886..1972
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2064..2091
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2110..2201
/gene="bla"
/label=AmpR promoter
CDS 2202..3062
/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(2420..2439)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3104..3559
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3191..3210)
/label=F1ori-R
/note="F1 origin, reverse primer"
rep_origin 3670..4258
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4159..4178
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4412..4429
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4594..4615
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4630..4660
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4668..4684
/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 4673..4695
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4692..4708
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4692..4708
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4724..4741)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4724..4740)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
61 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
121 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
181 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
241 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
301 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
361 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
421 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
481 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
541 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
601 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
661 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
721 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
781 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
841 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
901 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
961 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
1021 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
1081 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
1141 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
1201 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
1261 cgcaactctc tactgtttct ccatacccgt ttttttgggc tagcgattga aaacgatgca
1321 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
1381 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
1441 gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
1501 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
1561 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgtttt ggggaatata
1621 atctagcatt acgctaggga taacagggta atatcacgct ctagacatac ggcatgcaag
1681 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
1741 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
1801 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
1861 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
1921 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
1981 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
2041 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
2101 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
2161 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
2221 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
2281 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
2341 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
2401 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
2461 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
2521 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
2581 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
2641 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
2701 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
2761 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
2821 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
2881 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
2941 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
3001 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
3061 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
3121 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
3181 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
3241 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
3301 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
3361 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
3421 tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
3481 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
3541 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
3601 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
3661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
3721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
3781 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
3841 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
3901 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
3961 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
4021 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
4081 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
4141 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
4201 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
4261 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
4321 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
4381 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
4441 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtac
4501 gatttaaata ggcctgactc actataggga gaccggaatt ccctggcacg acaggtttcc
4561 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggg
4621 accccgggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
4681 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
4741 aacgtcgtga ctggtaaaac ccgggcgtta cccaacttaa tcgccttgca gcacatcccc
4801 ctttcgccag caggcgtaat aaggaaagga tccatgtact atttgaaaaa cacaaacttt
4861 tggatgttcg gtttattctt tttcttttac ttttttatca tgggagccta cttcccgttt
4921 ttcccgattt ggctacatga tatcaaccat atcagcaaaa gtgatacggg tattattttt
4981 gccgctattt ctctgttctc gctattattc caaccgctgt ttggtctgct ttctgacaaa
5041 ctcggtctac gcaaatacct gctgtggatt attaccggca tgttagtgat gtttgcgccg
5101 ttctttattt ttatcttcgg gccactgctg cagtacaaca ttttagtagg atcgattgtt
5161 ggtggtattt atctaggctt tagttttaac gccggtgcgc cagcagtaga ggcatttatt
5221 gagaaagtca gccggcgcag taatttcgaa tttggtcgcg cgcggatgtt tggcagtgtt
5281 ggctgggcgc tggttgcctc gattgtcggg atcatgttca ccattaataa tcagtttgtt
5341 ttctggctgg gctctggcag ttgtctcatc ctcgccgttt tactcttttt cgccaaaacg
5401 gacgcgccct cgagtgccac ggttgccaat gcggtaggtg ccaaccattc ggcatttagc
5461 cttaagctgg cactggaact gttcagacag ccaaaactgt ggtttttgtc actgtatgtt
5521 attggcgttt cctccaccta cgatgttttt gaccaacagt ttgctaattt ctttacttcg
5581 ttctttgcta ccggtgaaca gggtacccgc gtatttggct acgtaacgac aatgggcgaa
5641 ttacttaacg cctcgattat gttctttgcg ccactgatca ttaatcgcat cggtgggaag
5701 aatgccctgc tgctggctgg cactattatg tctgtacgta ttattggctc atcgttcgcc
5761 acctcagcgc tggaagtggt tattctgaaa acgctgcata tgtttgaagt accgttcctg
5821 ctggtgggct cctttaaata tattactagt cagtttgaag tgcgtttttc agcgacgatt
5881 tatctggtca gtttcagctt ctttaagcaa ctggcgatga tttttatgtc tgtactggcg
5941 ggcaatatgt atgaaagcat aggtttccaa ggcgcttatc tggtgctggg tctggtggcg
6001 ctgggcttca ccttaatttc cgtgttcacg cttagcggcc cgggcccgct ttccctgctg
6061 cgtcgtcagg tgaatgaagt cgcttaaagg cc
//
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