| 质粒名称: | pAG306GPD-EGFP-ccdB |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 14308 |
| 存储实验室: | Susan Lindquist |
|
载体骨架基本信息 |
|
| 载体名称: | pRS306 |
| 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: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 酵母表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pAG306GPD-EGFP-ccdB was a gift from Susan Lindquist (Addgene plasmid # 14308 ; http://n2t.net/addgene:14308 ; RRID:Addgene_14308) |
| 参考文献部分: | -- |
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 pAG306GPD-EGFP-ccdB 7673 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAG306GPD-EGFP-ccdB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7673)
TITLE S cerevisiae Advanced Gateway Destination Vectors
REFERENCE 2 (bases 1 to 7673)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7673
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 195..410
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 411..1214
/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="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(1343..1798)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1480..1501)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1692..1711
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1928..1950
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1942..1959
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1943..1959
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1966..1985
/label=T7
/note="T7 promoter, forward primer"
promoter 1966..1984
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2009..2653
/gene="S. cerevisiae TDH3"
/label=GAP promoter
/note="promoter for glyceraldehyde-3-phosphate
dehydrogenase"
primer_bind 2547..2570
/label=GPDpro-F
/note="S. cerevisiae GPD promoter, forward primer"
primer_bind 2659..2675
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2659..2675
/label=pBluescriptSK
/note="For pBluescript vector"
CDS 2688..3404
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2733..2754)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2994..3013)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3341..3362
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 3429..3553
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 3578..3608
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3662..4321
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(3728..3747)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 4666..4971
/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 4850..4869
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(5012..5136)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
terminator 5143..5390
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(5159..5177)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
promoter complement(5413..5431)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5452..5468)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5452..5468)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5465..5487)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5476..5492
/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)."
promoter complement(5500..5530)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(5683..5700)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5854..6442)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6613..7473)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7236..7255
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7474..7578)
/gene="bla"
/label=AmpR promoter
primer_bind 7646..7664
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
1981 tagggcgaat tggagctctt cgagtttatc attatcaata ctcgccattt caaagaatac
2041 gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt agccttttaa
2101 ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat atataacacc
2161 gaaggtgcct gggtgaacag gtttattcct ggcatccact aaatataatg gagcccgctt
2221 tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt ttcttcacca
2281 accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg cacaaacagg
2341 caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa tgatgacaca
2401 aggcaattga cccacgcatg tatctatctc attttcttac accttctatt accttctgct
2461 ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga aattattccc
2521 ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg taaatctatt
2581 tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt aaaacaccag
2641 aacttagttt cgacggattc tagaactagt ggatcccccg ggacaaaatg gtgagcaagg
2701 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
2761 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
2821 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
2881 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
2941 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
3001 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
3061 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
3121 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
3181 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
3241 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
3301 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
3361 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggtgga ccaggtggtg
3421 gacatcaaac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
3481 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
3541 tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc
3601 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
3661 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
3721 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
3781 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
3841 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
3901 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
3961 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
4021 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
4081 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
4141 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
4201 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtctgtg atggcttcca
4261 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
4321 aacgccgcgt ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga
4381 tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat
4441 gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca
4501 tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc
4561 tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta
4621 ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt
4681 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
4741 gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
4801 gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
4861 accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac
4921 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
4981 tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca
5041 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
5101 ttacgtttct cgttcagctt tcttgtacaa agtggttcga tgtcatgtaa ttagttatgt
5161 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
5221 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
5281 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
5341 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc
5401 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
5461 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
5521 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc
5581 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
5641 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
5701 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
5761 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
5821 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga
5881 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
5941 ccaggcgttc ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
6001 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
6061 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
6121 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
6181 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
6241 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
6301 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
6361 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
6421 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
6481 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
6541 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
6601 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
6661 tcgttcatcc atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt
6721 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
6781 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
6841 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
6901 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
6961 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
7021 gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
7081 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
7141 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
7201 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
7261 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
7321 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
7381 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
7441 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
7501 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
7561 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
7621 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
