| 质粒名称: | pLG163 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 124365 |
| 存储实验室: | Christopher Voigt |
|
载体骨架基本信息 |
|
| 载体名称: | pJAB186 |
| 载体出品公司: | -- |
| 空载体大小: | 7880 bp |
| 完整质粒大小: | 8591 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | gfpmut2 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | Pspank |
| 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 | caaatccgccgctctagctaag |
| 3’测序引物 | ttcagcacgtgtcttgtagttccc |
|
文献引用方法 |
|
| 材料和方法部分: | pLG163 was a gift from Christopher Voigt (Addgene plasmid # 124365 ; http://n2t.net/addgene:124365 ; RRID:Addgene_124365) |
| 参考文献部分: | Resilient living materials built by printing bacterial spores. Gonzalez LM, Mukhitov N, Voigt CA. Nat Chem Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 10.1038/s41589-019-0412-5. 10.1038/s41589-019-0412-5 PubMed 31792444 |
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 pLG163 8583 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION IPTG inducible system (gfpmut2 with Ppsank) for expression in B.
subtilis.
ACCESSION .
VERSION .
KEYWORDS pLG163
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8583)
AUTHORS Gonzalez LM, Mukhitov N, Voigt CA
TITLE Resilient living materials built by printing bacterial spores.
JOURNAL Nat Chem Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi:
10.1038/s41589-019-0412-5.
PUBMED 31792444
REFERENCE 2 (bases 1 to 8583)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8583
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 90..678
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 579..598
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 4187..4273
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4292..4319
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 4399..4433
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
protein_bind 4537..4553
/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)."
CDS 4586..5299
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(4613..4641)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 5236..5256
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
CDS 5555..6637
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(5574..5593)
/label=LacI-R
/note="LacI, reverse primer"
primer_bind complement(7451..7469)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7537..7641
/gene="bla"
/label=AmpR promoter
CDS 7642..8502
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(7860..7879)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
61 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
661 gggcggagcc tatggaaaaa cgccagcaac gcggcccgac ctcgagctgg atacttcccg
721 tccgccaggg ggacatgccg gcgatgctga aggtcgcgcg cattcccgat gaagaggccg
781 gttaccgcct gtttgaggat atagtaatct ttctaaatag ctttggattg gaggagtatg
841 gccactaata ctaagttcag ctaataaaaa aatttgctaa agaactccag ctggatttca
901 ctgatgagaa tatcgtcgga gataaatata ataattccac ggactataga ctatactagt
961 atactccgtc tactgtacga tacacttccg ctcaggtcct tgtcctttaa cgaggattgt
1021 tacgtacgct aatggcgtca aaacaaagac tctagaccta ggccttaaga tctgatcata
1081 tgcatccgcg ggcccgggtt aacgcgtaat ccatggatca agagacagga tgaggatcgt
1141 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggtgccctga atgaactgca
1201 gaaagagctg gtagttggcg cactgttcga agaactgccg atgtccagta agattcttac
1261 tatgctggtt gaaccggatg ctggtaaagc tacttgggtt gctgcttcta cttatggtac
1321 cgatacaact actggtgagg aagttaaagg agctcttaaa gaaatccact tcagtaggtg
1381 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
1441 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
1501 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
1561 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
1621 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gactaagaaa
1681 atgccgtcaa atccgctcgc catgacttca ctaacgatgc ctttgaaaat cttcaagttc
1741 ttttctacta attcaaggcg tgtctcacca ggtttttggt ttgctccggc gcaaatgcag
1801 acaatatcag catccttgca gggtatgttt ctctttgatg tctttttgtt tgtgaagtat
1861 ttcacattta tattgtgcaa cacttcacaa acttttgcaa gagaaaagtt ttgtctgatt
1921 tatgaacaaa aaagaaacca tcattgatgg tttctttcgg taagtcccgt ctagccttgc
1981 cctcaatggg gaagagaacc gcttaagccc gagtcattat ataaaccatt tagcacgtaa
2041 tcaaagccag gctgattctg accgggcact tgggcgctgc cattattaaa aatcactttt
2101 gcgttggttg tatccgtgtc cgcaggcagc gtcagcgtgt aaattccgtc tgcattttta
2161 gtcattggtt ttccaggcca agatccggtc aattcaatta ctcggctccc atcatgttta
2221 tagatataag catttacctg gctccaatga ttcggatttt gatagccgat ggttttggcc
2281 gacgctggat ctcttttaac aaaactgtat ttctcggtcc tcgttacacc atcactgttc
2341 gttcctttta acatgatggt gtatgttttg ccaaatggat ctccttttcc gattgtgaat
2401 tgatctccat ccttaaacgc cgtctctggt ccattattga tttgataaac ggcttttgtt
2461 gtattcgcat ctgcacgcaa ggtaatcgtc agttgatcat tgaaagaatg tgttacacct
2521 gttttgtaat tctcaaggaa aacatgaggc gcttttgcaa tatcatcagg ataaagcaca
2581 gctacagacc tggcattgat cgtgcctgtc agtttaccat cgttcacttg aaatgaaccc
2641 gctccagctt tattgtcata cctgccatca ggcaattttg ttgccgtatt gatagagaca
2701 gaggatgaac ctgcatttgc cagcacaacg ccatgtgagc cgcgctgatt cataaatatc
2761 tggttgtttc cattcgggtt cgagagttcc tcaggctgtc cagccatcac attgtgaaat
2821 ctattgaccg cagtgatagc ctgatcttca aataaagcac tcccgcgatc gcctatttgg
2881 cttttccccg ggaacctcac accatttccg cctccctcag gtctggaaaa gaaaagaggc
2941 gtactgcctg aacgagaagc tatcaccgcc cagcctaaac ggattccccc tatgcaaggg
3001 tttattgttt tctaaaatct gattaccaat tagaatgaat atttcccaaa tattaaataa
3061 taaaacaaaa aaattgaaaa aagtgtttcc accatttttt caattttttt ataatttttt
3121 taatctgtta tttaaatagt ttatagttaa atttacattt tcattagtcc attcaatatt
3181 ctctccaaga taactacgaa ctgctaacaa aattctctcc ctatgttcta atggagaaga
3241 ttcagccact gcatttcccg caatatcttt tggtatgatt ttacccgtgt ccatagttaa
3301 aatcatacgg cataaagtta atatagagtt ggtttcatca tcctgataat tatctattaa
3361 ttcctctgac gaatccataa tggctcttct cacatcagaa aatggaatat caggtagtaa
3421 ttcctctaag tcataatttc cgtatattct tttatttttt cgttttgctt ggtaaagcat
3481 tatggttaaa tctgaattta attccttctg aggaatgtat ccttgttcat aaagctcttg
3541 taaccattct ccataaataa attcttgttt gggaggatga ttccacggta ccatttcttg
3601 ctgaataata attgttaatt caatatatcg taagttgctt ttatctccta ttttttttga
3661 aataggtcta attttttgta taagtatttc tttactttga tctgtcaatg gttcagatac
3721 gacgactaaa aagtcaagat cactatttgg ttttagtcca ctctcaactc ctgatccaaa
3781 catgtaagta ccaataaggt tattttttaa atgtttccga agtatttttt tcactttatt
3841 aatttgttcg tatgtattca aatatatcct cctcactatt ttgattagta cctattttat
3901 atccatagtt gttaattaaa taaacttaat ttagtttatt tatagatttc attggcttct
3961 aaatttttta tctagataat aattatttta gttaatttta ttctagatta tatatgatat
4021 gatctttcat ttccataaaa ctaaagtaag tgtaaaccta ttcattgttt taaaaatatc
4081 tcttgccagt cacgttacgt tattagttat agttattata acatgtattc acgaacgaaa
4141 atcgccattc gccagctgca ggaattcgac ctctagcttg aggcatcaaa taaaacgaaa
4201 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
4261 gagtaggaca aatccgccgc tctagctaag cagaaggcca tcctgacgga tggccttttt
4321 gcgtttctac aaactcttgt taactctaga gctgcctgcc gcgtttcggt gatgaagatc
4381 ttcccgatga ttaattaatt cagaacgctc ggttgccgcc gggcgttttt tatgcagcaa
4441 tggcaagaac gttgctcgag ggtaaatgtg agcactcaca attcattttg caaaagttgt
4501 tgactttatc tacaaggtgt ggcataatgt gtgtaattgt gagcggataa caattaagct
4561 ttcggctgaa aggtggtgaa ctactatgaa aggagaagaa cttttcactg gagttgtccc
4621 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
4681 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
4741 acctgttcca tggccaacac ttgtcactac tttcgcgtat ggtcttcaat gctttgcgag
4801 atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt
4861 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa
4921 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
4981 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat
5041 ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga
5101 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
5161 ccttttacca gacaaccatt acctgtccac acaatctaag ctttcgaaag atcccaacga
5221 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
5281 ggatgaacta tacaaataag ctagccgcat gcaagctaat tcggtggaaa cgaggtcatc
5341 atttccttcc gaaaaaacgg ttgcatttaa atcttacata tgtaatactt tcaaagacta
5401 catttgtaag atttgatgtt tgagtcggct gaaagatcgt acgtaccaat tattgtttcg
5461 tgattgttca agccataaca ctgtagggat agtggaaaga gtgcttcatc tggttacgat
5521 caatcaaata ttcaaacgga gggagacgat tttgatgaaa ccagtaacgt tatacgatgt
5581 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
5641 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
5701 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
5761 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
5821 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaaac
5881 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
5941 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
6001 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
6061 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
6121 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
6181 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
6241 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
6301 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
6361 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
6421 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
6481 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
6541 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
6601 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
6661 aggatcctga gcgccggtcg ctaccattac cagttggtct ggtgtcaaaa ataataataa
6721 ccgggcaggc catgtctgcc cgtatttcgc gtaaggaaat ccattatgta ctatttaatt
6781 ctgcgtgaca tcccatcgat cagaccagtt tttaatttgt gtgtttccat gtgtccagtt
6841 tggaatactc ttaacctcat tggaaatcgc ggcataatca ctggtggtat gattgatgac
6901 cgcgtcaaca atgaccttta tgccatattc ttcagcggct gcacacattt ctttaaattc
6961 ttgttcagta cctaagtaac ggttgccaat ttgatacgat gtcggctgat acagccagta
7021 ccagttcgac atgcttttat ctccttgatt cccttccttt acttggttaa tcggagatgt
7081 ctgaatggct gtatatcctg catcatgaat atccttcata ttgtgtttta acgtattgaa
7141 cgaccaattc catgcatgaa gaatggttcc gcttttgatc gacggtgctg taagctcatt
7201 cgatttgttc gccgtttcag cactcgcagc cgccggtcct gccagaacca aatgaaacag
7261 caataaaaat ccagcgaata acggcagtaa agaggttttg aatcgttttg caaacattct
7321 tgacactcct tatttgattt tttgaagact tacttcggag tcaaaaatcc ctcttacttc
7381 attcttccgc ttcctccttt caaaccgatg tgaagactgg agaattttgt taattcttga
7441 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
7501 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
7561 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
7621 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
7681 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
7741 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
7801 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
7861 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
7921 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
7981 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
8041 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
8101 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
8161 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
8221 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
8281 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
8341 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
8401 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
8461 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
8521 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
8581 atc
//
