| 质粒名称: | pRE57I-GNAc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170637 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC57mini |
| 完整质粒大小: | 6420bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | Mini-Tn6677, GNAc cassette, lacI |
| 物种来源: | S. cerevisiae (budding yeast) |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pRE57I-GNAc was a gift from Sheng Yang (Addgene plasmid # 170637 ; http://n2t.net/addgene:170637 ; RRID:Addgene_170637) |
| 参考文献部分: | Programming Cells by Multicopy Chromosomal Integration Using CRISPR-Associated Transposases. Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, Yang S. CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018. 10.1089/crispr.2021.0018 PubMed 34152213 |
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 pRE57I-GNAc 6420 bp ds-DNA circular SYN 04-AUG-2021
DEFINITION DNA donor for transposition.
ACCESSION .
VERSION .
KEYWORDS pRE57I-GNAc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6420)
AUTHORS Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J,
Yang S
TITLE Programming Cells by Multicopy Chromosomal Integration Using
CRISPR-Associated Transposases.
JOURNAL CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018.
PUBMED 34152213
REFERENCE 2 (bases 1 to 6420)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 4, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6420
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 233..251
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 252..276
/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)."
RBS 291..313
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
promoter 2170..2188
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2189..2213
/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)."
RBS 2228..2250
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
terminator 2816..2864
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
terminator 2926..2974
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
primer_bind complement(3043..3065)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin complement(3504..4092)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3584..3603)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4263..5123)
/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 4886..4905
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5124..5228)
/gene="bla"
/label=AmpR promoter
CDS complement(5259..6341)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 6303..6322
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(6342..6419)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
ORIGIN
1 ccaatgatgc gaaaaaaccc cgccgaagcg gggttttttg cgaaaaaaat agggataaca
61 gggtaatgtt taaactgttg atacaaccat aaaatgataa ttacacccat aaattgataa
121 ttatcacacc cacgggaatt ccatggtcga cccagcaacc gcacctgtgg cgccggtgat
181 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
241 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
301 tttaagaagg agatatacca tgtgtggaat tgttggcgcg atcgcgcaac gtgatgtagc
361 agaaatcctt cttgaaggtt tacgtcgtct ggaataccgc ggatatgact ctgccggtct
421 ggccgttgtt gatacagaag gtcatatgac ccgcctgcgt cgcctcggta aagtccagat
481 gctggcacag gcagcggaag aacatcctct gcatggcggc actggtattg ctcacactcg
541 ctgggcgacc cacggtgaac cttcagaagt gaatgcgcat ccgcatgttt ctgaacacat
601 tgtggtggtg cataacggca tcatcgaaaa ccatgaaccg ctgcgtgaag agctaaaagc
661 gcgtggctat accttcgttt ctgaaaccga caccgaagtg attgcccatc tggtgaactg
721 ggagctgaaa caaggcggga ctctgcgtga ggccgttctg cgtgctatcc cgcagctgcg
781 tggtgcgtac ggtacagtga tcatggactc ccgtcacccg gataccctgc tggcggcacg
841 ttctggtagt ccgctggtga ttggcctggg gatgggcgaa aactttatcg cttctgacca
901 gctggcgctg ttgccggtga cccgtcgctt tatcttcctt gaagagggcg atattgcgga
961 aatcactcgc cgttcggtaa acatcttcga taaaactggc gcggaagtaa aacgtcagga
1021 tatcgaatcc aatctgcaat atgacgcggg cgataaaggc atttactgtc actacatgca
1081 gaaagagatc tacgaacagc cgaacgcgat caaaaacacc cttaccggac gcatcagcca
1141 cggtcaggtt gatttaagcg agctgggacc gaacgccgac gaactgctgt cgaaggttga
1201 gcatattcag atcctcgcct gtggtacttc ttataactcc ggtatggttt cccgctactg
1261 gtttgaatcg ctagcaggta ttccgtgcga cgtcgaaatc gcctctgaat tccgctatcg
1321 caaatctgcc gtgcgtcgta acagcctgat gatcaccttg tcacagtctg gcgaaaccgc
1381 ggataccctg gctggcctgc gtctgtcgaa agagctgggt taccttggtt cactggcaat
1441 ctgtaacgtt ccgggttctt ctctggtgcg cgaatccgat ctggcgctaa tgaccaacgc
1501 gggtacagaa atcggcgtgg catccactaa agcattcacc actcagttaa ctgtgctgtt
1561 gatgctggtg gcgaagctgt ctcgcctgaa aggtctggat gcctccattg aacatgacat
1621 cgtgcatggt ctgcaggcgc tgccgagccg tattgagcag atgctgtctc aggacaaacg
1681 cattgaagcg ctggcagaag atttctctga caaacatcac gcgctgttcc tgagccgtgg
1741 cgatcagtac ccaatcgcgc tggaaggcgc attgaagttg aaagagatct cttacattca
1801 cgctgaagcc tacgctgctg gcgaactgaa acacggtccg ctggcgctaa ttgatgccga
1861 tatgccggtt attgttgttg caccgaacaa cgaattgctg gaaaaactga aatccaacat
1921 tgaagaagtt cgcgcgcgtg gcggtcagtt gtatgtcttc gccgatcagg atgcgggttt
1981 tgtaagtagc gataacatgc acatcatcga gatgccgcat gtggaagagg tgattgcacc
2041 gatcttctac accgttccgc tgcagctgct ggcttaccat gtcgcgctga tcaaaggcac
2101 cgacgttgac cagccgcgta acctggcaaa atcggttacg gttgagtaag gatctcgatc
2161 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
2221 aaataatttt gtttaacttt aagaaggaga tataccatga gcttacccga tggattttat
2281 ataaggcgaa tggaagaggg agatttggaa caggtcactg agacgctaaa ggttttgacc
2341 accgtgggca ctattacccc cgaatccttc agcaaactca taaaatactg gaatgaagcc
2401 acagtatgga atgataacga agataaaaaa ataatgcaat ataaccccat ggtgattgtg
2461 gacaagcgca ccgagacggt tgccgctacg gggaatatca tcatcgaaag aaagatcatt
2521 catgaactgg ggctatgtgg ccacatcgag gacattgcag taaactccaa gtatcagggc
2581 caaggtttgg gcaagctctt gattgatcaa ttggtaacta tcggctttga ctacggttgt
2641 tataagatta ttttagattg cgatgagaaa aatgtcaaat tctatgaaaa atgtgggttt
2701 agcaacgcag gcgtggaaat gcaaattaga aaatagctcg agttaattaa actagtgagc
2761 tcggtacccg gggatcctct agaggtcgaa attcacctcg aaagcaagct gataaaccga
2821 tacaattaaa ggctcctttt ggagcctttt tttttggaga ttttcaacgt gaaaaaatta
2881 ttattcgcaa ttccaagcta attcacctcg aaagcaagct gataaaccga tacaattaaa
2941 ggctcctttt ggagcctttt tttttggaga ttttcaacgt gaaaaaatta ttattcgcaa
3001 ttccaagctc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
3061 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
3121 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
3181 cggagtgtat gggctcgatc ccctcggatc caattggtcc ttactgcagt agttttgctg
3241 aaatactcga ttcacaaaaa tatcaactta tggttgtttt gtgagatatc aatatatggt
3301 tgttttgtgg ttaagttgct gattataaat aattattaaa tatcacttta tggttgcatc
3361 aacagttaac attaccctgt tatccctaaa aaaaaaaccc cgcccctgac agggcggggt
3421 ttttttttta atatcggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
3481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
3541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
3601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
3661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
3721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
3781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
3841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
3901 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
3961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
4141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4261 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4321 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4381 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
4441 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
4501 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
4561 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
4621 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
4681 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
4741 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
4801 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
4861 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
4921 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
4981 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5041 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
5101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
5161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5221 gttccgcgca catttccccg aaaagtgcca cctgacgttc actgcccgct ttccagtcgg
5281 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
5341 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
5401 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
5461 cgaaaatcct gtttgatggt ggttgacggc gggatataac atgagctgtc ttcggtatcg
5521 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
5581 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
5641 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
5701 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
5761 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
5821 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
5881 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
5941 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
6001 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
6061 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
6121 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
6181 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
6241 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
6301 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
6361 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcc
//
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