| 质粒名称: | FDP-P10B2-A75-RZ-URA3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 130874 |
| 存储实验室: | Chang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | ColE1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Expression of mKate, Leu2, and Ura3 on orthogonal plasmid after integration onto p1 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mKate |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | FDP-P10B2-A75-RZ-URA3 was a gift from Chang Liu (Addgene plasmid # 130874 ; http://n2t.net/addgene:130874 ; RRID:Addgene_130874) |
| 参考文献部分: | Tunable Expression Systems for Orthogonal DNA Replication. Zhong Z, Ravikumar A, Liu CC. ACS Synth Biol. 2018 Dec 21;7(12):2930-2934. doi: 10.1021/acssynbio.8b00400. Epub 2018 Nov 16. 10.1021/acssynbio.8b00400 PubMed 30408954 |
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 FDP-P10B2-A75-RZ-URA3 6063 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION Integration cassette for OrthoRep containing mKate driven by
promoter 10B2 with a hardcoded polyA tail.
ACCESSION .
VERSION .
KEYWORDS FDP-P10B2-A75-RZ-URA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6063)
AUTHORS Zhong Z, Ravikumar A, Liu CC
TITLE Tunable Expression Systems for Orthogonal DNA Replication.
JOURNAL ACS Synth Biol. 2018 Dec 21;7(12):2930-2934. doi:
10.1021/acssynbio.8b00400. Epub 2018 Nov 16.
PUBMED 30408954
REFERENCE 2 (bases 1 to 6063)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6063
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1527..2225
/codon_start=1
/product="monomeric far-red fluorescent protein (Shcherbo
et al., 2009)"
/label=mKate2
/note="mammalian codon-optimized"
/translation="MHVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRI
KAVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVL
TATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMA
LKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVAR
YCDLPSKLGHR"
misc_RNA 2313..2364
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
terminator 2372..2536
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(2426..2450)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
CDS complement(2627..3430)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MHKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
CDS 3719..3910
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 4007..4146
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4062..4081
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4161..4178)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4332..4920)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4412..4431)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 4951..4990
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
promoter complement(5855..5946)
/gene="bla"
/label=AmpR promoter
terminator 6013..6060
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 6056..11
/label=T7 Term
/note="T7 terminator, reverse primer"
ORIGIN
1 ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc
61 ttctagaagt actatcaaaa tctgttattc tattgccttg aggatctttg aaacaccaat
121 ctgagtctga ggctacaatt tttcctttat aaccttcaac actatttaga gcttctttca
181 tttggttgta tctttcttgt cttatttctt ccatattaaa ttttatatct cttatatgaa
241 agtttttata ataattataa aatgtctgtt cctaagaaga tcgtcgtttt gccaggtgac
301 cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt
361 cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct
421 acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga tgccgttttg
481 ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta
541 ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc
601 gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt
661 gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat
721 ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg
781 gccgctttca tggccctata acatgagcca ccattgccta tttggtcctt ggataaagct
841 aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa
901 ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag
961 aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc
1021 gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct
1081 ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat
1141 ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa
1201 ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg
1261 gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat
1321 gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat
1381 ttgtacataa actttataaa tgaaattcat aatagaaacg ccgcgggatg acctatacat
1441 aggaagatct atagaaacaa aaagattaat aactttcaaa tatcagaaaa atgtagaaac
1501 atgtgataag ctcatagaca tgtaaaatgc atgtgagcga gctgattaag gagaacatgc
1561 acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg
1621 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggcggtc gagggcggcc
1681 ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc aaaaccttca
1741 tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccccgag ggcttcacat
1801 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
1861 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg
1921 gccctgtgat gcagaagaaa acactcggct gggaggcctc caccgagacc ctgtaccccg
1981 ctgacggcgg cctggaaggc agagccgaca tggccctgaa gctcgtgggc gggggccacc
2041 tgatctgcaa cttgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc
2101 ccggcgtcta ctatgtggac agaagactgg aaagaatcaa ggaggccgac aaagagacct
2161 acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc
2221 acagaggatc ctcatgaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
2281 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aacctgtcac cggatgtgtt ttccggtctg
2341 atgagtccgt gaggacgaaa cagggaattg ccgaatttct tatgatttat gatttttatt
2401 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
2461 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
2521 tagcatgagg tcgctcaagc ttgagctcgg gtaataactg atataattaa attgaagctc
2581 taatttgtga gtttagtata cctgcattta cttataatac agttttttag ttttgctggc
2641 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
2701 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
2761 ccacatcatc cacggttcta tactgttgac ccaatgcgtc acccttgtca tctaaaccca
2821 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
2881 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
2941 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
3001 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
3061 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagaatact
3121 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
3181 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
3241 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
3301 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
3361 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
3421 ctttatgcat tttataatta ttataaaaac tttcatatat tttaaattaa taatgtaatt
3481 catatattat agtactctcg agggtgccaa cacctacatc tgtattaacg aagcgctggc
3541 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
3601 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
3661 ctcgtttcat cggtatcatt accccatgaa cagaaatccc ccttacacgg aggcatcagt
3721 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
3781 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
3841 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
3901 aaacctctga cacatgcagc tcccgcgtca cagcttgtct gtaagcggat gccgggagca
3961 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
4021 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
4081 actgagagtg caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
4141 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
4201 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
4261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
4321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
4381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
4441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
4501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
4561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
4621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
4681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
4741 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
4801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
4861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
4921 gaagatcctt tgatcttttc taccggtccg tccaaaaaaa aaggctccaa aaggagcctt
4981 taattgtatc ggtttaccaa tgcttaatca gtgaagcccc tatttcagca atctgtctat
5041 ttcgttcgtc catagttgcc tgactacccg tcgtgtagat aactacgata cgggaaggct
5101 taccatctgg tccaagtgcg gcaatgatac cccgtgaacc acgctctcct gccccagatt
5161 tatcagcaat aaaccagcca gcaggaagtg ccgaacgcag tagtggtcct gcaactttat
5221 ccgcttcaag ccagtctatt aattgttgcc gtgaagccag agtaagtagt tcgccagtta
5281 atagtttgcg taatgttgtt gccattgccg caggcattgt ggtgtctcgc tcgtcgtttg
5341 gtatggcttc attcagttct ggttcccaac ggtcaaggcg agttacatgg tctcccatat
5401 tgtgcaaaaa agcggttaat tccttcggtc caccgatggt tgtcagaagt aagttcgccg
5461 cagtgttatc actcatcgtt atggcggcac tacataattc tcttaccgtc attccgtccg
5521 taaggtgctt ttctgtcact ggggagtatt caaccaagtc attctgcgaa tagtgtattc
5581 gtcgtccgag ttgctcttgc cccgcatcta cacgggataa tactgctcca catagcagaa
5641 ctttgaaagt gctcatcatt ggaaaacgct cttcagggcg aaaactctca aggattttac
5701 cgctattcaa gtccagttct atgtatccca ctcgtgcccc caattggtcc tctgcgtctt
5761 ttactttcac gagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
5821 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
5881 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
5941 aacaaatagg ggaattaaaa aaaagcccgc tcattaggcg ggctactagt cggatatagt
6001 tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag
6061 tta
//
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