| 质粒名称: | pBS-ldhGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 27170 |
| 存储实验室: | Michela Lizier |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript II KS(+) |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2931 bp |
| 完整质粒大小: | -- |
| 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: | lactate dehydrogenase promoter region fused with modified green fluorescent protein 5,Lactococcus lactis ldhL promoter,synthetic construct mgfp5 |
| 物种来源: | Lactobacillus acidophilus and synthetic construct |
| 插入大小: | 1001 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII (未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | M13 Reverse |
| 3' 测序引物 | M13 Forward |
|
文献引用方法 |
|
| 材料和方法部分: | pBS-ldhGFP was a gift from Michela Lizier (Addgene plasmid # 27170 ; http://n2t.net/addgene:27170 ; RRID:Addgene_27170) |
| 参考文献部分: | Comparison of expression vectors in Lactobacillus reuteri strains. Lizier M, Sarra PG, Cauda R, Lucchini F. FEMS Microbiol Lett. 2010 Jul 1. 308(1):8-15. 10.1111/j.1574-6968.2010.01978.x PubMed 20455948 |
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载体基本信息
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载体简介
载体序列
LOCUS pBS-ldhGFP 3932 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBS-ldhGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3932)
AUTHORS Lizier M, Sarra PG, Cauda R, Lucchini F
TITLE Comparison of expression vectors in Lactobacillus reuteri strains.
JOURNAL FEMS Microbiol Lett. 2010 Jul 1. 308(1):8-15.
PUBMED 20455948
REFERENCE 2 (bases 1 to 3932)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3932
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 6..722
/codon_start=1
/gene="mgfp5"
/product="GFP with folding enhancement mutations"
/label=mgfp5
/note="suitable for expression in plants due to removal of
a cryptic intron"
/translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
NFKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(36..64)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 659..679
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind complement(724..740)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(724..740)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(772..791)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(773..791)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(801..818)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(801..817)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(810..832)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 959..1414
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1046..1065)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1256..1277
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1440..1544
/gene="bla"
/label=AmpR promoter
CDS 1545..2405
/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 complement(1763..1782)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2576..3164
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3065..3084
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3318..3335
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3452..3473
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3488..3518
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3526..3542
/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)."
primer_bind 3531..3553
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3550..3566
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3550..3566
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 3585..3605
/label=T3
/note="T3 promoter, forward primer"
promoter 3587..3605
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 3634..3650
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 3635..3651
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 aattcatgag taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
61 atggtgatgt taatgggtac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
121 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa
181 cacttgtcac tactctcact tatggtgttc aatgcttttc aagatatcca gatcatatga
241 agcggcacga cttcttcaag agcgccatgc ctgagggata cgtgcaggag aggaccatct
301 tcttcaagga cgacgggaac tacaagacac gtgctgaagt caagtttgag ggagacaccc
361 tcgtcaacag gatcgagctt aagggaatcg atttcaagga ggacggaaac atcctcggcc
421 acaagttgga atacaactac aactcccaca acgtatacat catggccgac aagcaaaaga
481 acggcatcaa agccaacttc aagacccgcc acaacatcga agacggcggc gtgcaactcg
541 ctgatcatta tcaacaaaat actccaattg gcgatgaccc tgtcctttta ccagacaacc
601 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
661 tccttcttga gtttgtaacg gctgctggga ttacacatgg catggatgaa ctatacaaat
721 aaggatccac tagttctaga gcggccgcca ccgcggtgga gctccaattc gccctatagt
781 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
841 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
901 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
961 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
1021 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
1081 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
1141 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
1201 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
1261 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
1321 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
1381 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
1441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
1501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
1561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
1621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
1681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
1741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
1801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
1861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
1921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
1981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
2041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
2101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
2161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
2221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
2281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
2341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
2401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
2461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
2521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
2581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
2641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
2701 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
2761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
2821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
2881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
2941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
3001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
3061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
3121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
3181 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
3241 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
3301 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
3361 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
3421 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
3481 cccaggcttt acactttatg ctcccggctc gtatgttgtg tggaattgtg agcggataac
3541 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
3601 aaagggaaca aaagctgggt accgggcccc ccctcgaggt cgacggtatc gataagcttt
3661 ttagtccaat gcccttcaaa tttatcttta tttaaagtca aacctttttt atcaaggtag
3721 tgatccattg tcatgaaacc agtattatgt ttggtcttat catatttttg acccggatta
3781 cccaaacctg caattatctt catcttattt acccctcatt aataataatc tcaactataa
3841 tagcacaaac acaaaataat aattttatta atgctcttca acatggtata attttctttg
3901 ttaaaattat cactaataaa aaaggagact tg
//
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