| 质粒名称: | Dual opposing T7 promoters reporter plasmid |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 156457 |
| 存储实验室: | Matthew D. Shoulders |
|
载体骨架基本信息 |
|
| 载体名称: | BAC |
| 载体出品公司: | -- |
| 空载体大小: | 5862 bp |
| 完整质粒大小: | 7059 bp |
| 载体修饰: | 用氨苄青霉素抗性标记物代替氯霉素抗性标记物 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | NeoR/KanR |
| 物种来源: | Tn5 |
| 插入大小: | 804 bp |
| 基因突变: | ATG start codon mutated to ACG |
| GenBank编号 | -- |
| 启动子: | Em7 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制酶 |
| 5’克隆位点: | HindIII(未破坏) |
| 3’克隆位点: | PacI (未破坏) |
| 5’测序引物 | GTGCACCCAACTGATCTTCA |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Dual opposing T7 promoters reporter plasmid was a gift from Matthew D. Shoulders (Addgene plasmid # 156457 ; http://n2t.net/addgene:156457 ; RRID:Addgene_156457) |
| 参考文献部分: | A Processive Protein Chimera Introduces Mutations across Defined DNA Regions In Vivo. Moore CL, Papa LJ 3rd, Shoulders MD. J Am Chem Soc. 2018 Sep 19;140(37):11560-11564. doi: 10.1021/jacs.8b04001. Epub 2018 Jul 18. 10.1021/jacs.8b04001 PubMed 29991261 |
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 Dual opposing T7 promoters reporter plasmid 7059 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION A mutational reporter plasmid that can gain Kanamycin resistance
upon mutation. Contains two T7 promoters flanking the kanamycin
resistance gene..
ACCESSION .
VERSION .
KEYWORDS Dual opposing T7 promoters reporter plasmid
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7059)
AUTHORS Moore CL, Papa LJ 3rd, Shoulders MD
TITLE A Processive Protein Chimera Introduces Mutations across Defined DNA
Regions In Vivo.
JOURNAL J Am Chem Soc. 2018 Sep 19;140(37):11560-11564. doi:
10.1021/jacs.8b04001. Epub 2018 Jul 18.
PUBMED 29991261
REFERENCE 2 (bases 1 to 7059)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7059
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(9..27)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 308..781
/gene="sopC"
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
CDS complement(854..1825)
/codon_start=1
/gene="sopB"
/product="partitioning protein for the bacterial F plasmid"
/label=sopB
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
CDS complement(1825..3000)
/codon_start=1
/gene="sopA"
/product="partitioning protein for the bacterial F plasmid"
/label=sopA
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
misc_feature 3326..3576
/gene="incC"
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS complement(3579..4334)
/codon_start=1
/gene="repE"
/product="replication initiation protein for the bacterial
F plasmid"
/label=repE
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
rep_origin 4425..4644
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS complement(4870..5730)
/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 5493..5512
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5731..5835)
/gene="bla"
/label=AmpR promoter
promoter 5944..5962
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 6006..6053
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6075..6875
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
HQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6135..6154)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6745..6764
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
promoter complement(6954..6972)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
61 ttagacgtca ggtggcactt ttcggggaaa tgtgcccggt taacgtgccg gcacggcctg
121 ggtaaccagg tattttgtcc acataaccgt gcgcaaaatg ttgtggataa gcaggacaca
181 gcagcaatcc acagcaggca tacaaccgca caccgaggtt actccgttct acaggttacg
241 acgacatgtc aatacttgcc cttgacaggc attgatggaa tcgtagtctc acgctgatag
301 tctgatcgac aatacaagtg ggaccgtggt cccagaccga taatcagacc gacaacacga
361 gtgggatcgt ggtcccagac taataatcag accgacgata cgagtgggac cgtggtccca
421 gactaataat cagaccgacg atacgagtgg gaccgtggtt ccagactaat aatcagaccg
481 acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac gagtgggacc
541 atggtcccag actaataatc agaccgacga tacgagtggg accgtggtcc cagtctgatt
601 atcagaccga cgatacgagt gggaccgtgg tcccagacta ataatcagac cgacgatacg
661 agtgggaccg tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc
721 agtctgatta tcagaccgac gatacaagtg gaacagtggg cccagagaga atattcaggc
781 cagttatgct ttctggcctg taacaaagga cattaagtaa agacagataa acgtagacta
841 aaacgtggtc gcatcagggt gctggctttt caagttcctt aagaatggcc tcaattttct
901 ctatacactc agttggaaca cgagacctgt ccaggttaag caccatttta tcgcccttat
961 acaatactgt cgctccagga gcaaactgat gtcgtgagct taaactagtt cttgatgcag
1021 atgacgtttt aagcacagaa gttaaaagag tgataacttc ttcagcttca aatatcaccc
1081 cagctttttt ctgctcatga aggttagatg cctgctgctt aagtaattcc tctttatctg
1141 taaaggcttt ttgaagtgca tcacctgacc gggcagatag ttcaccgggg tgagaaaaaa
1201 gagcaacaac tgatttaggc aatttggcgg tgttgataca gcgggtaata atcttacgtg
1261 aaatattttc cgcatcagcc agcgcagaaa tatttccagc aaattcattc tgcaatcggc
1321 ttgcataacg ctgaccacgt tcataagcac ttgttgggcg ataatcgtta cccaatctgg
1381 ataatgcagc catctgctca tcatccagct cgccaaccag aacacgataa tcactttcgg
1441 taagtgcagc agctttacga cggcgactcc catcggcaat ttctatgaca ccagatactc
1501 ttcgaccgaa cgccggtgtc tgttgaccag tcagtagaaa agaagggatg agatcatcca
1561 gtgcgtcctc agtaagcagc tcctggtcac gttcattacc tgaccatacc cgagaggtct
1621 tctcaacact atcaccccgg agcacttcaa gagtaaactt cacatcccga ccacatacag
1681 gcaaagtaat ggcattaccg cgagccatta ctcctacgcg cgcaattaac gaatccacca
1741 tcggggcagc tggtgtcgat aacgaagtat cttcaaccgg ttgagtattg agcgtatgtt
1801 ttggaataac aggcgcacgc ttcattatct aatctcccag cgtggtttaa tcagacgatc
1861 gaaaatttca ttgcagacag gttcccaaat agaaagagca tttctccagg caccagttga
1921 agagcgttga tcaatggcct gttcaaaaac agttctcatc cggatctgac ctttaccaac
1981 ttcatccgtt tcacgtacaa cattttttag aaccatgctt ccccaggcat cccgaatttg
2041 ctcctccatc cacggggact gagagccatt actattgctg tatttggtaa gcaaaatacg
2101 tacatcaggc tcgaaccctt taagatcaac gttcttgagc agatcacgaa gcatatcgaa
2161 aaactgcagt gcggaggtgt agtcaaacaa ctcagcaggc gtgggaacaa tcagcacatc
2221 agcagcacat acgacattaa tcgtgccgat acccaggtta ggcgcgctgt caataactat
2281 gacatcatag tcatgagcaa cagtttcaat ggccagtcgg agcatcaggt gtggatcggt
2341 gggcagttta ccttcatcaa atttgcccat taactcagtt tcaatacggt gcagagccag
2401 acaggaagga ataatgtcaa gccccggcca gcaagtgggc tttattgcat aagtgacatc
2461 gtccttttcc ccaagataga aaggcaggag agtgtcttct gcatgaatat gaagatctgg
2521 tacccatccg tgatacattg aggctgttcc ctgggggtcg ttaccttcca cgagcaaaac
2581 acgtagcccc ttcagagcca gatcctgagc aagatgaaca gaaactgagg ttttgtaaac
2641 gccaccttta tgggcagcaa ccccgatcac cggtggaaat acgtcttcag cacgtcgcaa
2701 tcgcgtacca aacacatcac gcatatgatt aatttgttca attgtataac caacacgttg
2761 ctcaacccgt cctcgaattt ccatatccgg gtgcggtagt cgccctgctt tctcggcatc
2821 tctgatagcc tgagaagaaa ccccaactaa atccgctgct tcacctattc tccagcgccg
2881 ggttattttc ctcgcttccg ggctgtcatc attaaactgt gcaatggcga tagccttcgt
2941 catttcatga ccagcgttta tgcactggtt aagtgtttcc atgagtttca ttctgaacat
3001 cctttaatca ttgctttgcg tttttttatt aaatcttgca atttactgca aagcaacaac
3061 aaaatcgcaa agtcatcaaa aaaccgcaaa gttgtttaaa ataagagcaa cactacaaaa
3121 ggagataaga agagcacata cctcagtcac ttattatcac tagcgctcgc cgcagccgtg
3181 taaccgagca tagcgagcga actggcgagg aagcaaagaa gaactgttct gtcagatagc
3241 tcttacgctc agcgcaagaa gaaatatcca ccgtgggaaa aactccaggt agaggtacac
3301 acgcggatag ccaattcaga gtaataaact gtgataatca accctcatca atgatgacga
3361 actaaccccc gatatcaggt cacatgacga agggaaagag aaggaaatca actgtgacaa
3421 actgccctca aatttggctt ccttaaaaat tacagttcaa aaagtatgag aaaatccatg
3481 caggctgaag gaaacagcaa aactgtgaca aattaccctc agtaggtcag aacaaatgtg
3541 acgaaccacc ctcaaatctg tgacagataa ccctcagact atcctgtcgt catggaagtg
3601 atatcgcgga aggaaaatac gatatgagtc gtctggcggc ctttcttttt ctcaatgtat
3661 gagaggcgca ttggagttct gctgttgatc tcattaacac agacctgcag gaagcggcgg
3721 cggaagtcag gcatacgctg gtaactttga ggcagctggt aacgctctat gatccagtcg
3781 attttcagag agacgatgcc tgagccatcc ggcttacgat actgacacag ggattcgtat
3841 aaacgcatgg catacggatt ggtgatttct tttgtttcac taagccgaaa ctgcgtaaac
3901 cggttctgta acccgataaa gaagggaatg agatatgggt tgatatgtac actgtaaagc
3961 cctctggatg gactgtgcgc acgtttgata aaccaaggaa aagattcata gcctttttca
4021 tcgccggcat cctcttcagg gcgataaaaa accacttcct tccccgcgaa actcttcaat
4081 gcctgccgta tatccttact ggcttccgca gaggtcaatc cgaatatttc agcatattta
4141 gcaacatgga tctcgcagat accgtcatgt tcctgtaggg tgccatcaga ttttctgatc
4201 tggtcaacga acagatacag catacgtttt tgatcccggg agagactata tgccgcctca
4261 gtgaggtcgt ttgactggac gattcgcggg ctatttttac gtttcttgtg attgataacc
4321 gctgtttccg ccatgacaga tccatgtgaa gtgtgacaag tttttagatt gtcacactaa
4381 ataaaaaaga gtcaataagc agggataact ttgtgaaaaa acagcttctt ctgagggcaa
4441 tttgtcacag ggttaagggc aatttgtcac agacaggact gtcatttgag ggtgatttgt
4501 cacactgaaa gggcaatttg tcacaacacc ttctctagaa ccagcatgga taaaggccta
4561 caaggcgctc taaaaaagaa gatctaaaaa ctataaaaaa aataattata aaaatatccc
4621 cgtggataag tggataaccc caagggaagt tttttcaggc atcgtgtgta agcagaatat
4681 ataagtgctg ttccctggtg cttcctcgct cactcgagct acggggtctg acgctcagtg
4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct ggcggccgcc tttcagcaaa
5881 aaaccccgcg agacccccga agaggccccg cggggttatg ctaggtcgac ggagctcgaa
5941 ttctaatacg actcactata gggagaccca agctggcttg acaattaatc atcggctcga
6001 agcttgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
6061 ggaactaaac cacgggatcg gccattgaac aagatggatt gcacgcaggt tctccggccg
6121 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
6181 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
6241 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
6301 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
6361 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
6421 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
6481 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
6541 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
6601 tcaaggcgcg catgcccgac ggcgatgatc tcgtcgtgac ccatggcgat gcctgcttgc
6661 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
6721 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
6781 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
6841 tcgccttcta tcgccttctt gacgagttct tctgagggga tcaattctct agagcttaat
6901 taacgcagcc tgaatggcga atagaagttt aaacgctagc cagcttgggt ctccctatag
6961 tgagtcgtat tatcgagctc cgtcgaccta gcataacccc gcggggcctc ttcgggggtc
7021 tcgcggggtt ttttgctgaa agggatccaa ttcttgaag
//
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