| 质粒名称: | pSNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 135192 |
| 存储实验室: | Hongqing Fang |
|
载体骨架基本信息 |
|
| 载体名称: | pKOBEG |
| 完整质粒大小: | 7456 bp |
| 载体类型: | Bacterial Expression, Synthetic Biology |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | I-SceI lamba Red |
| 物种来源: | Synthetic |
| 基因突变: | 无 |
| 启动子: | araBAD |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI (not destroyed) |
| 3’克隆位点: | HindIII (not destroyed) |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pSNA was a gift from Hongqing Fang (Addgene plasmid # 135192 ; http://n2t.net/addgene:135192 ; RRID:Addgene_135192) |
| 参考文献部分: | An electroporation-free method based on Red recombineering for markerless deletion and genomic replacement in the Escherichia coli DH1 genome. Wei Y, Deng P, Mohsin A, Yang Y, Zhou H, Guo M, Fang H. PLoS One. 2017 Oct 24;12(10):e0186891. doi: 10.1371/journal.pone.0186891. eCollection 2017. 10.1371/journal.pone.0186891 PubMed |
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 pSNA 7456 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Arabinose inducible expression of lamba-Red recombinase proteins and
I-SceI. Contains ampicillin resistance cassette.
ACCESSION .
VERSION .
KEYWORDS pSNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7456)
AUTHORS Wei Y, Deng P, Mohsin A, Yang Y, Zhou H, Guo M, Fang H
TITLE An electroporation-free method based on Red recombineering for
markerless deletion and genomic replacement in the Escherichia coli
DH1 genome.
JOURNAL PLoS One. 2017 Oct 24;12(10):e0186891. doi:
10.1371/journal.pone.0186891. eCollection 2017.
PUBMED 29065183
REFERENCE 2 (bases 1 to 7456)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7456
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..285
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 208..227
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 1074..1490
/codon_start=1
/gene="gam"
/product="inhibitor of the host RecBCD nuclease in the
lambda Red system"
/label=Gam
/note="protects the double-stranded DNA substrate for
recombination"
/translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
CDS 1496..2281
/codon_start=1
/gene="bet"
/product="single-stranded DNA binding recombinase in the
lambda Red system"
/label=Beta
/note="mediates pairing of a single-stranded DNA with a
complementary target"
/translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
CDS 2278..2958
/codon_start=1
/gene="exo"
/product="5' to 3' double-stranded DNA exonuclease in the
lambda Red system"
/label=Exo
/note="creates 3' single-stranded DNA tails"
/translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
EQWR"
CDS complement(3456..4406)
/codon_start=1
/gene="rep101 encoding an A56V mutation"
/product="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/label=Rep101(Ts)
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 4454..4676
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS complement(5395..6255)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6018..6037
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6256..6360)
/gene="bla"
/label=AmpR promoter
CDS complement(6552..7430)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
301 ctagcaggag ggtacctata tgcatatgaa aatcatcaaa aaaaaccagg taatgaacct
361 cggtccgaac tctaaactgc tgaaagaata caaatcccag ctgatcgaac tgaacatcga
421 acagttcgaa gcaggtatcg gtctgatcct gggtgatgct tacatccgtt ctcgtgatga
481 aggtaaaacc tactgtatgc agttcgagtg gaaaaacaaa gcatacatgg accacgtatg
541 tctgctgtac gatcagtggg tactgtcccc gccgcacaaa aaagaacgtg ttaaccacct
601 gggtaacctg gtaatcacct ggggcgccca gactttcaaa caccaagcat tcaacaaact
661 ggctaacctg ttcatcgtta acaacaaaaa aaccatcccg aacaacctgg ttgaaaacta
721 cctgaccccg atgtctctgg catactggtt catggatgat ggtggtaaat gggattacaa
781 caaaaactct accaacaaat cgatcgtact gaacacccag tctttcactt tcgaagaagt
841 agaatacctg gttaagggtc tgcgtaacaa attccaactg aactgttacg taaaaatcaa
901 caaaaacaaa ccgatcatct acatcgattc tatgtcttac ctgatcttct acaacctgat
961 caaaccgtac ctgatcccgc agatgatgta caaactgccg aacactatct cctccgaaac
1021 tttcctgaaa taagctagaa gatgtttcgt gaagccgtcg acgcttataa aaaatggata
1081 ttaatactga aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa
1141 tcagcccggc atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg
1201 cttattacat tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg
1261 cccgtgaaga gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc
1321 tgtttgaatc gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta
1381 cccgtgcgtt tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca
1441 tggttgaaac cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag
1501 tactgcactc gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga
1561 cccacaggaa ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc
1621 gcagttcatc gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga
1681 aatttacgcc tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg
1741 gtcccgcatc atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga
1801 atcctgtaca tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg
1861 gatggatgaa tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc
1921 gtggcagtcg catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct
1981 ggccttcgga tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac
2041 tgcatacact gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat
2101 gcaggagatt aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc
2161 gctctgttcc cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga
2221 agcagtaaaa gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg
2281 acaccggaca ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggaa
2341 gatgcgtggc acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata
2401 gcaaaacccc gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg
2461 cttgctgagg tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga
2521 aaacagtacg agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact
2581 gaatccccga tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta
2641 tgcagtgacg gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag
2701 ttccggctcg gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc
2761 atgtgggtga cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt
2821 gaaggcctgc attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag
2881 atcgtgccgg agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt
2941 ggggagcaat ggcgatgacg catcctcacg ataataagct tcctgctgaa catcaaaggc
3001 aagaaaacat ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa
3061 taaaaacgca aaagaaaatg ccgatattga ctaccggaag cagtgtgacc gtgtgcttct
3121 caaatgcctg attcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt
3181 gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta
3241 catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa
3301 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
3361 acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg
3421 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
3481 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
3541 tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
3601 gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg
3661 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
3721 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
3781 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
3841 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
3901 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
3961 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
4021 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
4081 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
4141 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
4201 acaccataag cattttccct actgatgttc atcatctgaa cgtattggtt ataagtgaac
4261 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
4321 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
4381 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
4441 taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt
4501 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
4561 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
4621 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
4681 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
4741 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
4801 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
4861 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt
4921 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg
4981 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct
5041 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg
5101 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtcgggga
5161 tcgacgctct cccttatgcg actcctgcac ctttcgtctt cgaataaata cctgtgacgg
5221 aagatcactt cgcagaataa ataaatcctg gtctgacgct cagtggaacg aaaactcacg
5281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
5341 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
5401 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
5461 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
5521 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
5581 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
5641 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
5701 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
5761 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
5821 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
5881 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
5941 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
6001 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
6061 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
6121 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
6181 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
6241 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
6301 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
6361 cacatttccc cgaaaagtgt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
6421 tattgctggt ctcggtaccc ggggatcctc tagagtcgac ctgcataatg tgcctgtcaa
6481 atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga
6541 ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
6601 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga
6661 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
6721 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
6781 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
6841 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
6901 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
6961 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
7021 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
7081 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
7141 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
7201 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
7261 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
7321 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
7381 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
7441 ctcccgccat tcagag
//
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