| 质粒名称: | pMP2463 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 107774 |
| 存储实验室: | Herman Spaink |
|
载体骨架基本信息 |
|
| 载体名称: | pBBR1-mcs5 |
| 载体出品公司: | -- |
| 载体抗性: | 庆大霉素 |
| 空载体大小: | 4768 bp |
| 完整质粒大小: | 10368 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 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: | pLacZ-EGFP in pBBR1-mcs5 |
| 物种来源: | Aequorea victoria (Jellyfish) |
| 插入大小: | 5600 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | pLacZ |
| 标签/融合蛋白: | pLacZ EGFP（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Bam HI（销毁后未知） |
| 3' 克隆位点: | Xba I（销毁后未知） |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pMP2463 was a gift from Herman Spaink (Addgene plasmid # 107774 ; http://n2t.net/addgene:107774 ; RRID:Addgene_107774) |
| 参考文献部分: | Use of green fluorescent protein color variants expressed on stable broad-host-range vectors to visualize rhizobia interacting with plants. Stuurman N, Pacios Bras C, Schlaman HR, Wijfjes AH, Bloemberg G, Spaink HP. Mol Plant Microbe Interact. 2000 Nov;13(11):1163-9. doi: 10.1094/MPMI.2000.13.11.1163. 10.1094/MPMI.2000.13.11.1163 PubMed 11059482 |
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 pMP2463 5510 bp ds-DNA circular SYN 07-AUG-2018
DEFINITION Expression of EGFP.
ACCESSION .
VERSION .
KEYWORDS pMP2463
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5510)
AUTHORS Stuurman N, Pacios Bras C, Schlaman HR, Wijfjes AH, Bloemberg G,
Spaink HP
TITLE Use of green fluorescent protein color variants expressed on stable
broad-host-range vectors to visualize rhizobia interacting with
plants.
JOURNAL Mol Plant Microbe Interact. 2000 Nov;13(11):1163-9. doi:
10.1094/MPMI.2000.13.11.1163.
PUBMED 11059482
REFERENCE 2 (bases 1 to 5510)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5510
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 5..27
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 24..40
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 24..40
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 59..79
/label=T3
/note="T3 promoter, forward primer"
promoter 61..79
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 108..124
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 109..125
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
regulatory 171..180
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 177..896
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(222..243)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(483..502)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 830..851
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(946..965)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(947..965)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(975..992)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(975..991)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(984..1006)
/label=M13/pUC Forward
/note="In lacZ gene"
CDS complement(1700..2362)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/label=pBBR1 Rep
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
rep_origin 2363..3134
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS complement(4416..4949)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(5138..5166)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 5436..5457
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5472..5502
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5510..16
/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)."
ORIGIN
1 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
61 aattaaccct cactaaaggg aacaaaagct gggtaccggg ccccccctcg aggtcgacgg
121 tatcgataag cttgatatcg aattcctgca gcccggggga tccaccggtc gccaccatgg
181 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
241 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
301 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
361 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
421 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
481 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
541 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
601 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
661 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
721 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
781 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
841 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
901 gccgcgactc tagagcggcc gccaccgcgg tggagctcca attcgcccta tagtgagtcg
961 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
1021 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
1081 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa
1141 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
1201 aataggccgt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
1261 tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga
1321 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt
1381 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
1441 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga
1501 tcatttattc tgcctcccag agcctgataa aaacggtgaa tccgttagcg aggtgccgcc
1561 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
1621 gacaaggtat agggcggcga ggcggctaca gccgatagtc tggaacagcg cacttacggg
1681 ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc ggctccaacg
1741 gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc
1801 gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg
1861 gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg
1921 tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa
1981 ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca
2041 cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga
2101 tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga
2161 tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca
2221 cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct
2281 catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga
2341 tttctctgga ctgcgtggcc atgcttatct ccatgcggta ggggtgccgc acggttgcgg
2401 caccatgcgc aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa
2461 agtggcagtc aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg
2521 caatgagctg ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc
2581 gccctatatc tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc
2641 ttcggcgcgg ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg
2701 cggccttcgg ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt
2761 gaggctcggg gggcaggcgg gcgggcttcg cccttcgact gcccccactc gcataggctt
2821 gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc
2881 gccttccact tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt
2941 ccccagagaa aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg
3001 tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc
3061 aggcgcagcc tgtccatcag cttgtccagc agggttgtcc acgggccgag cgaagcgagc
3121 cagccggtgg ccgctcgcgg ccatcgtcca catatccacg ggctggcaag ggagcgcagc
3181 gaccgcgcag ggcgaagccc ggagagcaag cccgtagggc gccgcagccg ccgtaggcgg
3241 tcacgacttt gcgaagcaaa gtctagtgag tatactcaag cattgagtgg cccgccggag
3301 gcaccgcctt gcgctgcccc cgtcgagccg gttggacacc aaaagggagg ggcaggcatg
3361 gcggcatacg cgatcatgcg atgcaagaag ctggcgaaaa tgggcaacgt ggcggccagt
3421 ctcaagcacg cctaccgcga gcgcgagacg cccaacgctg acgccagcag gacgccagag
3481 aacgagcact gggcggccag cagcaccgat gaagcgatgg gccgactgcg cgagttgctg
3541 ccagagaagc ggcgcaagga cgctgtgttg gcggtcgagt acgtcatgac ggccagcccg
3601 gaatggtgga agtcggccag ccaagaacag caggcggcgt tcttcgagaa ggcgcacaag
3661 tggctggcgg acaagtacgg ggcggatcgc atcgtgacgg ccagcatcca ccgtgacgaa
3721 accagcccgc acatgaccgc gttcgtggtg ccgctgacgc aggacggcag gctgtcggcc
3781 aaggagttca tcggcaacaa agcgcagatg acccgcgacc agaccacgtt tgcggccgct
3841 gtggccgatc tagggctgca acggggcatc gagggcagca aggcacgtca cacgcgcatt
3901 caggcgttct acgaggccct ggagcggcca ccagtgggcc acgtcaccat cagcccgcaa
3961 gcggtcgagc cacgcgccta tgcaccgcag ggattggccg aaaagctggg aatctcaaag
4021 cgcgttgaga cgccggaagc cgtggccgac cggctgacaa aagcggttcg gcaggggtat
4081 gagcctgccc tacaggccgc cgcaggagcg cgtgagatgc gcaagaaggc cgatcaagcc
4141 caagagacgg cccgagacct tcgggagcgc ctgaagcccg ttctggacgc cctggggccg
4201 ttgaatcggg atatgcaggc caaggccgcc gcgatcatca aggccgtggg cgaaaagctg
4261 ctgacggaac agcgggaagt ccagcgccag aaacaggccc agcgccagca ggaacgcggg
4321 cgcgcacatt tccccgaaaa gtgccacctg cggcgttgtg acaatttacc gaacaactcc
4381 gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata
4441 tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg
4501 tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg
4561 aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctctccgga gactgcgaga
4621 tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga
4681 gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc
4741 aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac
4801 tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta
4861 catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc
4921 tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
4981 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc
5041 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
5101 ggaccagttg cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta
5161 tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg cgtccatggg caaatattat
5221 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
5281 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt ttttatgcat gcgcccaata
5341 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
5401 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
5461 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
//
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