| 质粒名称: | pCN-PDC1-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 45326 |
| 存储实验室: | Brendan Cormack |
|
载体骨架基本信息 |
|
| 载体名称: | pBM16 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 6672 bp |
| 载体修饰: | 用于将PDC1pr-GFP终止子片段从pCU-PDC1-GFP移动到pBM16骨架的SacI/Acc65I消化物。 |
| 载体类型: | C.glabrata表达 |
| 筛选标记: | nourseothricin (NAT) |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | yeGFP |
| 物种来源: | Synthetic |
| 插入大小: | 717 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | PDC1 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未知是否破坏) |
| 3’克隆位点: | SalI (未知是否破坏) |
| 5’测序引物 | M13F |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCN-PDC1-GFP was a gift from Brendan Cormack (Addgene plasmid # 45326 ; http://n2t.net/addgene:45326 ; RRID:Addgene_45326) |
| 参考文献部分: | Expression plasmids for use in Candida glabrata. Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J, Cormack BP. G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908. 10.1534/g3.113.006908 PubMed 23934995 |
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 pCN-PDC1-GFP 6672 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCN-PDC1-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6672)
AUTHORS Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J,
Cormack BP
TITLE Expression plasmids for use in Candida glabrata.
JOURNAL G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.
PUBMED 23934995
REFERENCE 2 (bases 1 to 6672)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6672
/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 188..585
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 599..1171
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
terminator 1178..1425
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(1194..1212)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind 1610..1632
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1624..1641
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1625..1641
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2627..2643
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2627..2643
/label=pBluescriptSK
/note="For pBluescript vector"
CDS 2663..3379
/codon_start=1
/product="yeast-enhanced green fluorescent protein"
/label=yeGFP
/note="codons optimized for translation in S. cerevisiae
and C. albicans"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(3853..3869)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3853..3869)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3866..3888)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3877..3893
/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(3901..3931)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3946..3967
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4084..4101)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4255..4843)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4335..4354)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5014..5874)
/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 5637..5656
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5875..5979)
/gene="bla"
/label=AmpR promoter
primer_bind 6645..6663
/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 accataccat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
241 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt
301 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
361 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc
421 ttgaaaattt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
481 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
541 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagttttg gatctaaaat
601 gggcaccact cttgacgaca cggcttaccg ataccgcacc agtgtcccgg gggacgccga
661 ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg tcaccgccac
721 cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca aggtgttccc
781 cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgaccctg actcccggac
841 gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct cgtactccgg
901 ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg
961 ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagctgg gcgccgggca
1021 cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc ggcggatggg
1081 gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg acggcgagca
1141 ggcgctctac atgagcatgc cctgccccta agtcgagtca tgtaattagt tatgtcacgc
1201 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
1261 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
1321 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
1381 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg tatgcggtgt
1441 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
1501 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
1561 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
1621 cgttgtaaaa cgacggccag tgaattcgag ctcagcattt ttatacacgt tttacgtatt
1681 ttttttgcaa tacacataga tacgtacgta caaccctttc tattgtgaga actttgtaac
1741 actcttttgt agaccagtgt agtcagcaag catcagcaag catcagcaag catcagcaag
1801 catcagctag tctaaagcct ttagcaaagt caaaaaccaa cacccacaca ccgggatccc
1861 accttccccg agatgcctgt tcccgactcc cagctgctcc cacgacgggg aggaatcaga
1921 agacggagat agaaagagag cgttgaacgc gcgcgatgcc aatgagagat aaaaaaaaag
1981 aaataccaat cccacgacat attctaacgg gctccacaga caatgcacaa caatcgagat
2041 gggattgaat cacagagaga gagatcagaa aaaaaaggaa ctgcaagggt ccccacccca
2101 aactcccctc cgctatccac tacaatcgcc gcaccgacaa aatccattgt gtccgcaacg
2161 accagagcaa agagccaaaa acacaattga ataacaaaga aaagggcaga gagccactta
2221 gacaattacg taggaaccat atttttgcat aactttatgc aatttgccca aattcaagtg
2281 aagcgagaga agccagagag ctacagctga agagctacag ctgaagtgaa aagccaagaa
2341 tctcttagaa tataccgttt ggcttatttg aatcaaatat ttatgcgctt aaatcccata
2401 aaaaagcaat ttatacagat aagtctgcag aaagaaaaat tttcttcttg atacctgaac
2461 aaagaaatca aactcatcaa gaataaatca attcatgaaa aaaaaacata tataaaggac
2521 aacatggaat caagtttcaa taatttttag attgtacata aataaagaga ccagactaat
2581 acaactgtat aagctctaaa cattcaattg ccaaaaaaca ttaacatcta gaactagtgg
2641 atcccccggg ctgcaggaat tcatgtctaa aggtgaagaa ttattcactg gtgttgtccc
2701 aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg
2761 tgaaggtgat gctacttacg gtaaattgac cttaaaattt atttgtacta ctggtaaatt
2821 gccggttcca tggccaacct tagtcactac tttcggttat ggtgttcaat gttttgctag
2881 atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt
2941 tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa
3001 gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga
3061 tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg tttacatcat
3121 ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa attagacaca acattgaaga
3181 tggttctgtt caattagctg accattatca acaaaatact ccaattggtg atggtccagt
3241 cttgttacca gacaaccatt acttatccac tcaatctgcc ttatccaaag atccaaacga
3301 aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat
3361 ggatgaattg tacaaataac tgcaggtcga cctcgagaac acaacccaca gctaccacca
3421 tcaacaatat ttatatatat aacgtacaca tagaaatcac acaaacagag tatttattct
3481 taactacatg aactaccatc agaccgtctg ggccactata taatgtgcca ttcataaacg
3541 tgatcacttt acgtagcagg caaccccagg tgaaaatttt tcagcgagct gccagattgt
3601 caggtgaaaa actgaaaaaa acttctgggc gatgagcttg tggcgggaaa ttaagtatat
3661 aaagcagtta gtttcttctg cttcttgtgg ttctgggtat tcttgtaata tttggaagaa
3721 tagaaaatca agactactgc ctttcttttc atattataga ggaagaaata cgcacgaaca
3781 cgatatagag gtaaaggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt
3841 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
3901 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
3961 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
4021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
4681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
4801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
4861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
4921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
4981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
5041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
5101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
5161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
5221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
5281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
5341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
5401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
5461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
5521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
5581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
5641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
5701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
5761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
5821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
5881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
5941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
6001 acctgacgtc gggatctcta atgatgcaat ttttgttttg tacactaaga ttctatttcg
6061 gaactcttac ccatgtcatt atttacttta ttttcattat tgtaattttt taagctttta
6121 ttatgaatta tttaaataca attctttttg atcatgtgac agacatcact gaaagatgtg
6181 ctctgcgtta ttagtcagat tgaaatatac attttcatgt agatatgatg ttcttacttt
6241 aaatgttgtt ccatttgcta gcttaaatag acgatattat aaatttaagt attcactaaa
6301 aatctgaccc attaatgtta gaattagtat tattttttgt gatattacat ttttatattt
6361 tggatttcaa ccaatgctct gcgatatgat tcaacagaat aatatgtaca agctacttta
6421 attcatatat actaatagat tcacagatcc agtacgactg caagaattta cgttttgttt
6481 gaaatggtag tagctacact aaattaacaa tgatcgatta atccacatcc acagtgtgta
6541 atcatcctaa tgaaaacaag aataactaca aaaatgaata taaacttttt gcatgttagt
6601 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
6661 ggccctttcg tc
//
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