| 质粒名称: | pBFC0996 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 186695 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | R6K conditional origin, bla, RP4 oriT |
| 载体出品公司: | -- |
| 完整质粒大小: | 10679 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | EC100D pir+ |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | tnsA tnsB tnsC tnsD tniQ cas8-cas5 fusion cas7 cas6 crRNA Tn7 transposon 2xLguI Cargo Stuffer SbfI+AsiSI dual restriction site for donor plasmid removal during ET-Seq |
| 物种来源: | Vibrio cholerae |
|
文献引用方法 |
|
| 材料和方法部分: | pBFC0996 was a gift from Jennifer Doudna (Addgene plasmid # 186695 ; http://n2t.net/addgene:186695 ; RRID:Addgene_186695) |
| 参考文献部分: | Species- and site-specific genome editing in complex bacterial communities. Rubin BE, Diamond S, Cress BF, Crits-Christoph A, Lou YC, Borges AL, Shivram H, He C, Xu M, Zhou Z, Smith SJ, Rovinsky R, Smock DCJ, Tang K, Owens TK, Krishnappa N, Sachdeva R, Barrangou R, Deutschbauer AM, Banfield JF, Doudna JA. Nat Microbiol. 2022 Jan;7(1):34-47. doi: 10.1038/s41564-021-01014-7. Epub 2021 Dec 6. 10.1038/s41564-021-01014-7 PubMed 34873292 |
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 pBFC0996 10679 bp ds-DNA circular SYN 09-SEP-2022
DEFINITION VcDART under control of Lac promoter, Vc_2xBsaI_NT gRNA, 2xLguI
cargo stuffer, with ET-Seq (AsiSI+SbfI) restriction sites flanking
Tn7 Right End.
ACCESSION .
VERSION .
KEYWORDS pBFC0996
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10679)
AUTHORS Rubin BE, Diamond S, Cress BF, Crits-Christoph A, Lou YC, Borges AL,
Shivram H, He C, Xu M, Zhou Z, Smith SJ, Rovinsky R, Smock DCJ, Tang
K, Owens TK, Krishnappa N, Sachdeva R, Barrangou R, Deutschbauer AM,
Banfield JF, Doudna JA
TITLE Species- and site-specific genome editing in complex bacterial
communities.
JOURNAL Nat Microbiol. 2022 Jan;7(1):34-47. doi: 10.1038/s41564-021-01014-7.
Epub 2021 Dec 6.
PUBMED 34873292
REFERENCE 2 (bases 1 to 10679)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 9, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10679
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 310..698
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
promoter 848..878
/label=lac promoter
/note="promoter for the E. coli lac operon"
RBS 879..890
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2975..2986
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 9481..9500
/label=T7
/note="T7 promoter, forward primer"
promoter 9481..9499
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 9623..9643
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
oriT 9687..9795
/note="incP origin of transfer"
promoter 9988..10092
/gene="bla"
/label=AmpR promoter
CDS 10093..274
/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(10311..10330)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
61 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
121 tgagcgtggc tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
181 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
241 tgagataggt gcctcactga ttaagcattg gtaactgtca gacccggcca cgatgcgtcc
301 ggcgtagagg atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat
361 gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac
421 cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta
481 ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa
541 ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt ttaaggttta
601 acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa
661 gcaattttga gtgacacagg aacacttaac ggctgacatg ggcaagttta ctcatatata
721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
781 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgaca cctgcccgaa
841 ccatatcttt acactttatg cttccggctc gtatgttgaa agaggagaaa ggatcaatgg
901 cgacaagttt acctacgccc tcagcaatta cgacttcggc gttagagtat gcattccata
961 ctcccgctcg caatctaacg aaatctcgcg gaaaaaacat tcatcgttat gtcagtgtaa
1021 agatgagtaa gaggattacg gtagaatcta ctctagagtg tgatgcctgc tatcactttg
1081 attttgagcc aagtattgtt cgcttttgcg ctcaaccgat tcgattttta tattatctca
1141 atggtcagtc tcactcctat gttcctgact ttctagttca atttgatacc aacgagtttg
1201 ttctatatga agtaaagtca gcttatgcta agaacaaacc tgattttgat gttgaatggg
1261 aggcgaaagt aaaagcagca actgaactag ggctagaatt ggagcttgtt gaagagagtg
1321 atattaggga tacggttgta ttaaataatc ttaagcgcat gcatcgttat gcttcgaaag
1381 atgagctgaa taacgtacat aactctctct taaaaataat aaagtacaat ggcgcccaat
1441 ctgcaagatg cttgggagaa cagttgggtt taaaaggccg aactgtttta ccaattttgt
1501 gcgatttgct gtcaaggtgt ttactcgata cacgtttgga taagcctcta tctcttgaat
1561 ctcgatttga gttggccagt tatggctaag aaagggttct caagtttcca tagaaaagca
1621 gtctcttccc aagatacgct tgaatcgata gagcttgtct ctagcgctaa ttgtctagaa
1681 agtgttacgt atcaagatat atcagcattt cccgaaacaa ttgcggtaga gattaatttc
1741 cgattaagca ttcttcgctt tttagcgcga aagtgcgaaa ccattgtggc caaatcaatt
1801 gaaccacatc gtgtagagct acagcaaaac tatagtagaa aaatacccag tgcaataacg
1861 atatatcgat ggtggcttgc ttttcgaaaa tcagactaca accccattag cttagcacct
1921 aatatcaagg atagaggtaa tagagagaca aaagtgtcaa cagttgttga ttctattatg
1981 gaacaggcag ttgaaagagt tatatctgga cgaaaagtca atgttagctc tgcatataaa
2041 cgtgttcgac gaaaagttcg tcaatacaat ctaactcatg gaacgaaata cacgtatcct
2101 aagtacgaat ctgtaagaaa gcgagtaaaa aagaaaaccc catttgagtt attagccgca
2161 gggaaagggg agagagtagc taagagagag tttcgccgaa tgggaaaaaa gatcctcacg
2221 tctagcgtgc tagagagggt tgaaatagat cacactgtcg ttgacctttt tgcagtacac
2281 gaagagtatc gaatcccatt gggccgacct tggcttactc aattggttga ttgttacagt
2341 aaagctgtaa tcggttttta tttaggtttc gagcctccta gctatgtgtc ggtttccctt
2401 gcacttaaga atgcaataca acgcaaagat gacttaatct cctcgtatga atcgatcgag
2461 aatgaatggc tatgttatgg catcccagac ctactcgtaa ctgataatgg taaagagttt
2521 ttgtcgaaag catttgatca agcatgtgaa tcactattga tcaatgtgca tcaaaataaa
2581 gttgaaacgc ccgacaacaa acctcatgtt gaacgtaact acgggactat taatacttct
2641 ctgttagacg atttacctgg gaaatccttc agccagtacc ttcaaagaga agggtacgac
2701 tctgtgggag aagctaccct tacactcaat gagattagag aaatttactt aatttggttg
2761 gtggatattt atcataaaaa acccaatcag agaggcacta attgtcctaa tgttgcgtgg
2821 aaaaagggtt gtcaagaatg ggaaccagag gagttctctg gttctaaaga cgaattagac
2881 tttaaatttg ctattgttga ttacaaacaa cttactaaag tagggataac tgtctacaaa
2941 gaactgagtt atagcaatga ccgtttagct gaatatagag ggaagaaagg aaaccataaa
3001 gttcagttca agtataaccc tgagtgtatg gcagttattt gggtgttgga tgaggatatg
3061 aatgagtact ttacagttaa tgcgattgac tacgaatatg caagtagagt atcactttgg
3121 caacataaat ataacatgaa atatcaagca gaactaaatt cagcagaata tgatgaggac
3181 aaggaaattg atgcagaaat aaaaattgaa gaaatcgcag atcgttcaat tgttaagact
3241 aacaaaatca gagctcggag gcgtggcgct aggcatcaag agaatagcgc aagggctaag
3301 tcaatcagta atgcgaaccc ggcctcgata caaaaacatg aagatgaaat cgttagtgca
3361 gataatgacg attgggatat tgattatgtc tgagaaatcg tcaaatgagt gaaacgcgtg
3421 aggctcgtat atcaagagct aaaagggcat ttgtatccac accgagcgtt aggaaaatct
3481 tgagttacat ggatagatgt agagatctat cagacctaga gtctgagcct acatgcatga
3541 tggtctatgg tgcttcgggt gtaggtaaaa cgaccgtcat caagaaatac ttaaatcaaa
3601 acagaagaga gtccgaagcc gggggcgata taataccggt tttgcatatt gagttgccag
3661 acaatgcgaa gccagtagat gcagcaaggg aattgctggt tgaaatgggt gacccgctag
3721 cactttatga aactgactta gctagattga cgaaaagact gactgaatta atccctgcgg
3781 tcggcgtgaa gctgattatt atcgatgagt tccaacattt ggtggaagaa aggtcaaatc
3841 gggttcttac ccaagtaggt aattggctaa aaatgatact taacaaaacg aaatgtccaa
3901 ttgttatatt tggtatgcca tactcaaaag ttgtactgca agcaaactcg caacttcacg
3961 ggcgattttc cattcaggtt gaactgcgcc cctttagcta ccagggaggt agaggtgtat
4021 ttaaaacttt tttggaatac cttgataaag ccctaccttt tgaaaaacag gctggcttag
4081 ccaacgaaag tttgcagaaa aaattgtatg cattctctca gggaaacatg cgttcgttga
4141 gaaaccttat ttatcaagca tctatcgaag caattgataa tcagcatgaa acgataaccg
4201 aagaagattt cgtttttgca tcgaagttga catcgggcga taaacccaac tcatggaaaa
4261 atccttttga ggagggtgtt gaggtaacag aagatatgtt acgaccgcca ccaaaagata
4321 ttggttggga ggactatttg agacattcaa ccccgagagt gagtaaacca ggtagaaata
4381 aaaacttttt cgaataacct aggctgctgc caccgctgag caataaccat cttagtatat
4441 tagttaagta taagaaggag atatacatat gtttttgcaa agacctaaac cttacagcga
4501 tgaaagttta gaaagtttct ttatccgagt ggctaacaaa aatggctacg gtgatgtcca
4561 tcgcttccta gaagccacta aacgattcct tcaagacatt gaccataatg gctatcaaac
4621 ctttccgact gatataactc ggataaaccc atactcagct aaaaacagtt ccagcgcacg
4681 aactgcgtca ttcctgaaac ttgcacaatt gacatttaat gaaccgccag agctacttgg
4741 gttggcaatt aacagaacaa acatgaaata ctcgccgtca actagcgcgg ttgttagagg
4801 tgcagaagtc tttcctcgca gtttactacg gacgcactcc atcccctgct gtcctttgtg
4861 tctgcgagaa aatggctacg cctcctacct ttggcacttt caggggtacg aatactgcca
4921 cagccataac gtacctttaa ttaccacttg tagctgtggt aaggagtttg actaccgagt
4981 atctgggtta aagggcattt gctgcaaatg caaggagcct atcaccttaa ccagcaggga
5041 gaacggtcat gaggcagcgt gtactgtttc aaactggctt gctggccatg aatctaaacc
5101 tctgccaaat cttcctaaaa gctaccgatg gggtttagtt cattggtgga tgggtattaa
5161 agatagcgag ttcgatcact tttcgttcgt tcaatttttc tcaaactggc caaggtcatt
5221 ccactcgata atcgaagatg aagtagagtt caaccttgag catgctgttg tcagcacgtc
5281 tgaattacga ctaaaagatc ttcttggtcg attgtttttc ggttcaattc ggttacctga
5341 gcggaatctt caacacaata tcatccttgg tgagcttctc tgctatttag aaaatcgttt
5401 atggcaagac aagggattaa tcgccaacct caaaatgaac gcgttagagg cgactgtaat
5461 gttaaattgt agcctcgatc agattgcatc aatggttgaa caacgcatct tgaagccaaa
5521 tcgaaaaagc aagcccaaca gccctcttga tgttaccgat tatctatttc atttcggcga
5581 tattttctgt ctttggttag ctgagttcca aagcgatgag tttaaccgtt cgttttatgt
5641 gtcgaggtgg taaatgcaaa ctctgaaaga actaatcgca tccaatcctg acgacttaac
5701 aactgaactt aagagagcat ttcgtccact cacaccgcat attgcaattg atggtaatga
5761 acttgacgca ctgacgatat tagtcaattt aaccgataag actgatgatc agaaagacct
5821 gctcgatcga gccaaatgca agcaaaaact tcgagatgaa aaatggtggg ctagctgcat
5881 aaattgcgtt aattacagac aaagccataa cccaaaattc ccggatatac gttctgaagg
5941 cgtgattcga acccaagccc tgggtgaatt accgagtttc ctactctcgt cctcaaaaat
6001 cccaccatac cattggtcat atagccatga ttcaaaatac gtcaacaaaa gcgcattcct
6061 caccaatgag ttttgttggg atggtgagat ctcatgtttg ggtgagcttc ttaaagatgc
6121 agatcaccca ctttggaata ctttgaaaaa gttaggttgt tctcaaaaaa cttgcaaagc
6181 aatggcaaaa caactagctg atattactct cacgactatc aatgtcacac tcgcaccaaa
6241 ttacctgact caaatatctc tccccgatag tgatacatcc tacatttcac tctctcctgt
6301 tgcatcgcta tcgatgcaaa gccactttca tcagaggctc caagatgaaa atcggcatag
6361 cgcgataacg cggtttagcc gaactaccaa catgggggtc actgcgatga catgtggcgg
6421 cgcatttaga atgttaaagt ctggcgctaa gttttccagc ccccctcatc accgattaaa
6481 cagtaaacga agttggttga cgtcagagca tgttcagtca ttaaaacagt accagcgcct
6541 caacaaaagc ctcatacctg aaaactctcg gattgcactc cgtagaaaat acaaaatcga
6601 gcttcaaaat atggtcagat cttggtttgc aatgcaagac catacccttg attcgaacat
6661 acttatccaa catttgaatc atgacctatc ttacttagga gccacaaaac gttttgcata
6721 cgatccagcg atgaccaagc tctttactga gcttttgaaa cgagagttat caaattcaat
6781 caataatggt gagcaacaca ctaatggatc gtttttagtc ctaccgaata tcagggtttg
6841 tggcgcaaca gctttaagct ccccggtaac ggtggggatt ccatcactta cagctttctt
6901 tggcttcgtt cacgcatttg aacggaatat aaatcgcacc acctcatcgt ttcgtgttga
6961 atcctttgcg atatgcgtcc atcaactaca tgtcgagaag cgaggtttga cagcagagtt
7021 tgtggaaaaa ggcgacggga ctatatccgc tcccgcgacc cgggatgact ggcagtgtga
7081 tgtcgtattt agccttattt tgaacaccaa ctttgctcaa catattgacc aagatacgtt
7141 agttacatca ctaccaaagc gattggctcg gggttcagca aaaattgcga ttgatgactt
7201 taaacatatc aactcattct cgacattaga aacagcgatc gaatctctgc caatagaagc
7261 tggtaggtgg ttatcacttt acgcacagtc aaacaataat ctaagtgatc tattagcagc
7321 catgacagag gaccatcagc tcatggcaag ctgcgtcggt taccacttgt tagaagaacc
7381 caaagataaa ccaaactccc tcagaggtta caaacacgct atcgccgagt gcatcattgg
7441 actcattaac tcaatcacct ttagctcaga gactgatccc aacacaatct tttggtcgct
7501 aaagaactat caaaactacc tagtggtaca gccaaggagt atcaacgatg aaactaccga
7561 caaatctagc ctatgagcgc tctatcgacc catcagatgt ctgttttttt gtcgtctggc
7621 ccgatgatag aaaaacacct ttaacctaca attctcgtac tctgctcggg caaatggaag
7681 cggcatcatt agcctatgat gtctcaggtc aaccaataaa aagtgccacc gctgaggcgt
7741 tagctcaagg gaaccctcat caagttgatt tctgccacgt tccatacggt gcgagtcata
7801 ttgaatgcag tttctccgtt tcgttttctt ctgaactacg tcaaccatat aagtgtaact
7861 caagcaaagt taaacaaacg ctagtgcaat tagtcgagct ctacgaaacg aaaatcggct
7921 ggactgagct agcaacccga tatttgatga acatttgcaa cggtaaatgg ctgtggaaaa
7981 atacccgtaa agcctattgc tggaacattg tacttacacc ttggccttgg aacggggaaa
8041 aggttggatt tgaagatatt cgtactaact acacctcacg gcaagacttt aaaaataata
8101 aaaattggtc tgctatagtt gaaatgatca aaaccgcatt ttctagtact gatgggctgg
8161 cgatatttga agtcagggcc accttgcact tgccaacgaa tgctatggtg cggccaagcc
8221 aagttttcac agaaaaagaa agtggcagta aaagtaaatc taaaactcaa aacagtcgag
8281 tttttcagag tacaactatt gatggtgaac gatcgccaat actaggggcc tttaaaacgg
8341 gagcagctat tgcaaccatt gacgactggt atcctgaagc cactgagcca ctaagggtcg
8401 gacggtttgg ggttcatcgc gaagatgtca cttgctaccg tcatccgtct accggaaaag
8461 attttttctc gatattacaa caagcagagc actatattga agtgttgagc gccaacaaaa
8521 ctcccgctca agaaactatc aacgacatgc actttttaat ggctaacctg attaagggtg
8581 ggatgttcca gcataaagga gactgactgt gaaatggtat tataagacaa tcacctttct
8641 gccagagttg tgcaacaacg agtcactggc tgcaaagtgt ctccgcgttc tgcatggatt
8701 taactatcag tatgagacac gaaatatcgg cgtttcattt ccgctttggt gtgatgcaac
8761 ggttggaaaa aagatttcat ttgtcagcaa gaacaagata gaactcgact tactacttaa
8821 acaacactat ttcgtccaaa tggaacaact tcaatacttt catatatcca acactgttct
8881 cgtcccagaa gattgtacat acgtttcctt tagacgctgt caatctatag ataagctcac
8941 agcagcaggg ctggcaagga aaatcagacg cctggagaaa cgtgctctat ctagaggcga
9001 gcaatttgac ccatcatctt ttgctcaaaa agagcatact gcaatagcgc actaccactc
9061 acttggggag tccagcaaac agacgaaccg caactttcga ctcaatatca ggatgctctc
9121 ggagcaaccc cgtgagggga actcgatttt tagtagctat ggcttatcaa attcagaaaa
9181 ctcgtttcag cctgtaccct taatctgacc taggctgctg ccaccggtga actgccgagt
9241 aggtagctga taacgagacc tcgtttacct atcggtctcg tgaactgccg agtaggtagc
9301 tgataaccca aaccagatgt caacacagct actgttgatg caaccataaa gtgatattta
9361 ataattattt ataatcagca acttaaccac aaaacaacca tatattgata tctcacaaaa
9421 caaccataag ttgatatttt tcagaaagga agagcgagat ctcgagctct tcggctgctg
9481 taatacgact cactataggg gataagagac ggctcaacgc ccgtctcaca gcgcaatatc
9541 aatttatggg tgtgataatt atcaatttat gggtgtaatt atcattttat ggttgtatca
9601 acaaggagga ctttggcagg tgcagcacaa aaggaaactc accgcgatcg ccattactcg
9661 catcagcctg caggattctt gcttatcggc cagcctcgca gagcaggatt cccgttgagc
9721 accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact
9781 tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt
9841 tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg
9901 accccgaagc agggttatgc agcggaaaag aatgtcatga taataatggt ttcttagacg
9961 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
10021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
10081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
10141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
10201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
10261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
10321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
10381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
10441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
10501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
10561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
10621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaact
//
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