| 质粒名称: | pBWB162 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140634 |
| 存储实验室: | Keith Tyo |
|
载体骨架基本信息 |
|
| 载体名称: | pBAV1k |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 5013bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Kanamycin, 25 μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | mCherry |
| 插入大小: | 711bp |
|
文献引用方法 |
|
| 材料和方法部分: | pBWB162 was a gift from Keith Tyo (Addgene plasmid # 140634 ; http://n2t.net/addgene:140634 ; RRID:Addgene_140634) |
| 参考文献部分: | Development of a genetic toolset for the highly engineerable and metabolically versatile Acinetobacter baylyi ADP1. Biggs BW, Bedore SR, Arvay E, Huang S, Subramanian H, McIntyre EA, Duscent-Maitland CV, Neidle EL, Tyo KEJ. Nucleic Acids Res. 2020 Apr 4. pii: 5815822. doi: 10.1093/nar/gkaa167. 10.1093/nar/gkaa167 PubMed 32246719 |
买家导航
pBWB162载体质粒基本信息
pBWB162质粒图谱载体图谱和pBWB162载体序列质粒序列多克隆位点信息
pBWB162质粒载体简介
pBWB162质粒序列载体序列
LOCUS pBWB162 5013 bp DNA circular SYN 14-APR-2025
DEFINITION pBAV1k-lacI-Trc-mCherry base, lacI repression vector for ADP1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5013)
AUTHORS Biggs BW, Bedore SR, Arvay E, Huang S, Subramanian H, McIntyre EA,
Duscent-Maitland CV, Neidle EL, Tyo KEJ
TITLE Development of a genetic toolset for the highly engineerable and
metabolically versatile Acinetobacter baylyi ADP1.
JOURNAL Nucleic Acids Res. 2020 Apr 4. pii: 5815822. doi:
10.1093/nar/gkaa167.
PUBMED 32246719
REFERENCE 2 (bases 1 to 5013)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. 2020 Apr 4. pii: 5815822. doi: 10.1093/nar/gkaa167."
COMMENT Sequence Label: pBWB162
FEATURES Location/Qualifiers
source 1..5013
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 34..45
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
terminator complement(247..341)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(357..1151)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 1357..1376
/label=BioBrick prefix
/note="BioBrick prefix for parts that start with ""ATG"""
promoter 1403..1480
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 1481..2563
/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
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(1500..1519)
/label=LacI-R
/note="LacI, reverse primer"
promoter join(2795..2800,2801..2817,2818..2824)
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 2832..2848
/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 2837..2859
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 2869..3579
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(3014..3032)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3282..3301
/label=mCherry-F
/note="mCherry, forward primer"
terminator 3630..3716
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ccctcatact cccttgagcc tcctccaacc gaaatagaag ggcgctgcgc ttattatttc
61 attcagtcat cggctttcat aatctaacag acaacatctt cgctgcaaag ccacgctacg
121 ctcaagggct tttacgctac gataacgcct gttttaacga ttatgccgat aactaaacga
181 aataaacgct aaaacgtctc agaaacgatt ttgagacgtt ttaataaaaa atcgcctagt
241 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
301 atggagttct gaggtcatta ctggatctat caacgggagt ccaagcgagc tcggtactaa
361 aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt gatccccagt
421 aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat cgaccggacg
481 cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc aataaagcca
541 cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca
601 agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg atctttaaat
661 ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa
721 tccaattcgg ctaagcggcc gtctaagcta ttcgtatagg gacaatccga tatgtcgatg
781 gagtgaaaga gcctgatgca ctccgcatac agctcgatag tcttttcagg gctttgttca
841 tcttcatacc cttccgagca aaggacgcca tcggcctcac tcatgagcag attgctccag
901 ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc cagccatagc
961 atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct gtccgtcatt
1021 tttaaatata ggatttcatt ttctcccacc agcttatata ccttagcagg agacattcct
1081 tccgtatctt ttacgcagcg gtattcttcg atcagttttt tcaattccgg tgatattctc
1141 attttagcca tttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag
1201 ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac cttaaataca
1261 gaaaacagcc ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
1321 gattttgaaa ccacaattat gatagaattt gacgtcgaat tcgcggccgc ttctagaact
1381 atagctagca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc gcggtatggc
1441 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
1501 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
1561 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
1621 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
1681 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
1741 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
1801 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
1861 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
1921 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
1981 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
2041 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
2101 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
2161 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
2221 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
2281 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
2341 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
2401 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
2461 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
2521 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
2581 taagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg tgcaccaatg
2641 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
2701 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
2761 ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc cggctcgtat
2821 aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaccat ggtgagcaag
2881 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt tcacatggag
2941 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
3001 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
3061 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
3121 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
3181 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaagacgg cgagttcatc
3241 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
3301 actatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc gctgaagggc
3361 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
3421 acctacaagg ccaagaagcc cgtgcaactg cccggcgcgt acaacgtcaa catcaagttg
3481 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
3541 cgccactcca ccggcggcat ggacgagctg tataagtaag cctcagggcc cgatcgatgc
3601 cgccgcttaa ttaattaatc cagaggcatc aaataaaacg aaaggctcag tcgaaagact
3661 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
3721 cgccctagac ctagtgtcat tttatttccc ccgtttcagc atcaagaacc tttgcataac
3781 ttgctctata tccacactga taattgccct caaaccataa tctaaaggcg ctagagtttg
3841 ttgaaacaat atcttttaca tcattcgtat ttaaaattcc aaactccgct cccctaaggc
3901 gaataaaagc cattaaatct tttgtattta ccaaattata gtcatccact atatctaaga
3961 gtaaattctt caattctctt ttttggcttt catcaagtgt tatatagcgg tcaatatcaa
4021 aatcattaat gttcaaaata tcttttttgt cgtatatatg tttattctta gcaatagcgt
4081 cctttgattc atgagtcaaa tattcatatg aacctttgat ataatcaagt atctcaacat
4141 gagcaactga actattcccc aattttcgct taatcttgtt cctaacgctt tctattgtta
4201 caggatttcg tgcaatatat ataacgtgat agtgtggttt tttatagtgc tttccatttc
4261 gtataacatc actactattc catgtatctt tatctttttt ttcgtccata tcgtgtaaag
4321 gactgacagc catagatacg cccaaactct ctaatttttc cttccaatca ttaggaattg
4381 agtcaggata taataaaaat ccaaaatttc tagctttagt atttttaata gccatgatat
4441 aattacctta tcaaaaacaa gtagcgaaaa ctcgtatcct tctaaaaacg cgagctttcg
4501 cttanttttt ttgttctgat tcctttcttg catattcttc tatagctaac gccgcaaccg
4561 cagattttga aaaacctttt tgtttcgcca tatctgttaa ttttttatct tgctcttttg
4621 tcagagaaat cataactctt tttttcgatt ctgaaatcac catttaaaaa actccaatca
4681 aataatttta taaagttagt gtatcacttt gtaatcataa aaacaacaat aaagctactt
4741 aaatatagat ttataaaaaa cgttggcgaa aacgttggcg attcgttggc gattgaaaaa
4801 ccccttaaac ccttgagcca gttgggatag agcgtttttg gcacaaaaat tggcactcgg
4861 cacttaatgg ggggtcgtag tacggaagca aaattcgctt cctttccccc catttttttc
4921 caaattccag atttttttca aaaattttcc agcgctaccg ctcggcaaaa ttgcaagcaa
4981 tttttaaaat caaacccatg agggaatttc att
//
