| 质粒名称: | pAB035h |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79191 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | SC101 |
| 载体出品公司: | David Liu Lab |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2992 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Plac62 gIII, luxAB; Ptet rpoZ-HA4 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Plac62; Ptet |
| 标签/融合蛋白: | -- |
| 克隆方法: | Ligation Independent 克隆 |
| 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: | pAB035h was a gift from David Liu (Addgene plasmid # 79191 ; http://n2t.net/addgene:79191 ; RRID:Addgene_79191) |
| 参考文献部分: | Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance. Badran AH, Guzov VM, Huai Q, Kemp MM, Vishwanath P, Kain W, Nance AM, Evdokimov A, Moshiri F, Turner KH, Wang P, Malvar T, Liu DR. Nature. 2016 May 5;533(7601):58-63. doi: 10.1038/nature17938. Epub 2016 Apr 27. 10.1038/nature17938 PubMed 27120167 |
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 pAB035h 7341 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Directed Evolution of Protein-Protein Interactions.
ACCESSION .
VERSION .
KEYWORDS pAB035h
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7341)
AUTHORS Badran AH, Guzov VM, Huai Q, Kemp MM, Vishwanath P, Kain W, Nance
AM, Evdokimov A, Moshiri F, Turner KH, Wang P, Malvar T, Liu DR
TITLE Continuous evolution of Bacillus thuringiensis toxins overcomes
insect resistance.
JOURNAL Nature. 2016 May 5;533(7601):58-63. doi: 10.1038/nature17938. Epub
2016 Apr 27.
PUBMED 27120167
REFERENCE 2 (bases 1 to 7341)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7341
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(1..951)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 999..1221
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
promoter 1818..1848
/label=lac promoter
/note="promoter for the E. coli lac operon"
CDS 3162..4241
/codon_start=1
/gene="luxA"
/product="LuxA luciferase subunit"
/label=LuxA
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
CDS 4305..5285
/codon_start=1
/gene="luxB"
/product="LuxB luciferase subunit"
/label=LuxB
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
CDS complement(5696..5725)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(5735..6007)
/codon_start=1
/gene="E. coli rpoZ"
/product="DNA-directed RNA polymerase subunit omega"
/label=RNAP omega
/note="promotes RNA polymerase assembly"
/translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGRR"
promoter 6071..6125
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 6072..6090
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6102..6120
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 6126..6144
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS complement(6270..7130)
/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 6893..6912
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7131..7235)
/gene="bla"
/label=AmpR promoter
primer_bind 7303..7321
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
1681 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
1741 aagactgggc ctttcgtttt gctgaggaga cttagggacc ctacaacacg cacggtgtta
1801 cattaggcac cccgggcttt acactttatg cttccggctc gtatgttgtg tcgaccgacc
1861 tgcaggtgca gtaaggagga aaaaaaaatg aaaaaattat tattcgcaat tcctttagtt
1921 gttcctttct attctcactc cgctgaaact gttgaaagtt gtttagcaaa accccataca
1981 gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat
2041 gagggctgtc tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt
2101 tacggtacat gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag
2161 ggtggcggtt ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt
2221 gatacaccta ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt
2281 actgagcaaa accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact
2341 ttcatgtttc agaataatag gttccgaaat aggcaggggg cattaactgt ttatacgggc
2401 actgttactc aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca
2461 aaagccatgt atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc
2521 tttaatgagg atccattcgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct
2581 cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggtggc
2641 tctgagggtg gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct
2701 ggttccggtg attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa
2761 aatgccgatg aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact
2821 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
2881 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
2941 aattcacctt taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa
3001 tgtcgccctt ttgtctttgg cgctggtaaa ccttacgagt tcagtatcga ctgcgataag
3061 atcaacctgt tccgcggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
3121 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aatgaaattt ggaaactttt
3181 tgcttacata ccaacctccc caattttccc aaacagaggt aatgaaacgt ttggttaaat
3241 taggtcgcat ctctgaggag tgtggttttg ataccgtatg gttactggag catcatttca
3301 cggagtttgg tttgcttggt aacccttatg tcgctgctgc atatttactt ggcgcgacta
3361 aaaaattgaa tgtaggaact gccgctattg ttcttcccac agcccatcca gtacgccaac
3421 ttgaagatgt gaatttattg gatcaaatgt caaaaggacg atttcggttt ggtatttgcc
3481 gagggcttta caacaaggac tttcgcgtat tcggcacaga tatgaataac agtcgcgcct
3541 tagcggaatg ctggtacggg ctgataaaga atggcatgac agagggatat atggaagctg
3601 ataatgaaca tatcaagttc cataaggtaa aagtaaaccc cgcggcgtat agcagaggtg
3661 gcgcaccggt ttatgtggtg gctgaatcag cttcgacgac tgagtgggct gctcaatttg
3721 gcctaccgat gatattaagt tggattataa atactaacga aaagaaagca caacttgagc
3781 tttataatga agtggctcaa gaatatgggc acgatattca taatatcgac cattgcttat
3841 catatataac atctgtagat catgactcaa ttaaagcgaa agagatttgc cggaaatttc
3901 tggggcattg gtatgattct tatgtgaatg ctacgactat ttttgatgat tcagaccaaa
3961 caagaggtta tgatttcaat aaagggcagt ggcgtgactt tgtattaaaa ggacataaag
4021 atactaatcg ccgtattgat tacagttacg aaatcaatcc cgtgggaacg ccgcaggaat
4081 gtattgacat aattcaaaaa gacattgatg ctacaggaat atcaaatatt tgttgtggat
4141 ttgaagctaa tggaacagta gacgaaatta ttgcttccat gaagctcttc cagtctgatg
4201 tcatgccatt tcttaaagaa aaacaacgtt cgctattata ttatggcggt ggcggtagcg
4261 gcggtggcgg tagcggcggt ggcggtagcg gcggtggcgg tagcaaattt ggattgttct
4321 tccttaactt catcaattca acaactgttc aagaacagag tatagttcgc atgcaggaaa
4381 taacggagta tgttgataag ttgaattttg aacagatttt agtgtatgaa aatcattttt
4441 cagataatgg tgttgtcggc gctcctctga ctgtttctgg ttttctgctc ggtttaacag
4501 agaaaattaa aattggttca ttaaatcaca tcattacaac tcatcatcct gtccgcatag
4561 cggaggaagc ttgcttattg gatcagttaa gtgaagggag atttatttta gggtttagtg
4621 attgcgaaaa aaaagatgaa atgcattttt ttaatcgccc ggttgaatat caacagcaac
4681 tatttgaaga gtgttatgaa atcattaacg atgctttaac aacaggctat tgtaatccag
4741 ataacgattt ttatagcttc cctaaaatat ctgtaaatcc ccatgcttat acgccaggcg
4801 gacctcggaa atatgtaaca gcaaccagtc atcatattgt tgagtgggcg gccaaaaaag
4861 gtattcctct catctttaag tgggatgatt ctaatgatgt tagatatgaa tatgctgaaa
4921 gatataaagc cgttgcggat aaatatgacg ttgacctatc agagatagac catcagttaa
4981 tgatattagt taactataac gaagatagta ataaagctaa acaagagacg cgtgcattta
5041 ttagtgatta tgttcttgaa atgcacccta atgaaaattt cgaaaataaa cttgaagaaa
5101 taattgcaga aaacgctgtc ggaaattata cggagtgtat aactgcggct aagttggcaa
5161 ttgaaaagtg tggtgcgaaa agtgtattgc tgtcctttga accaatgaat gatttgatga
5221 gccaaaaaaa tgtaatcaat attgttgatg ataatattaa gaagtaccac acggaatata
5281 cctaaacgga gccaatgtac gcaaaaaacc ccgcttcagc ggggtttttt cgccaaaaaa
5341 aaccccgccc tgtcaggggc ggggtttttt tttagggccc tactgactgt tagcaggtgc
5401 ggtaattgat actaatcgga gaatacatga acatgtaacc cgcgctatct tcgccccatg
5461 cataaaccgt gatggtgtaa tccacacccg ggctcaggcc actaatcgtc gcggtagagc
5521 tactatacgg aaccgtaaat tcctgcaccg ggctgttacc gcccgtttca ccgtaggtga
5581 tacgatagta atacacagag ctactagaca tcggggcatc ccaagaaatc agcaggctcg
5641 tcggggttgc tgcaaccact tccagtttgg tcggaacgct actcacagag ctgcccaagt
5701 cctcttcaga aataagcttt tgttccgcgg ccgcacgacg accttcagca atagcggtaa
5761 cggcttgtaa ttcagcggct tcctgctctt gctgttcctg gcgttcgcga acgtcgagga
5821 tctggttgtt gatcagacct tcttcgattt cgcgcagcgc gattacagtg gttttatcgt
5881 tttcttccgg taccagcgga tcctttccgc ctacctgcat ctgacgagcg cgacgcgcgg
5941 cgaccagtac caggtcaaaa cggttaccaa ttttctctac agcgtcctga acagttacgc
6001 gtgccatttt tttttcctcc ttataagggg gtagggggtc tagatttttg tcgaactatt
6061 catttcactt ttctctatca ctgataggga gtggtaaaat aactctatca atgatagagt
6121 gtcaatctct atcactgata gggatcatct ggccattcga tggacttaat taacggcact
6181 gcgaaaaaac cccgccgaag cggggttttt tgcgaaatat aatgaccctc ttgataaccc
6241 aagagggcat tttttaatgc ccatggcgtt taccaatgct taatcagtga ggcacctatc
6301 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
6361 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
6421 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
6481 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
6541 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
6601 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
6661 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
6721 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
6781 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
6841 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
6901 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
6961 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
7021 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
7081 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
7141 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
7201 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
7261 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
7321 cccttaggcc aaataggccg t
//
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