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负20摄氏度
6month
pUC119
货源充足
LSM Bio
无
2ug
Search name
pUC119 Informaiton
Promoter: Lac/lac promoter
Replicon: ColE1 ori, F1 ori
Plasmid size: 3162bp
Plasmid tagging: LacZ
Prokaryotic resistance: ampicillin Amp
Clonal strain: DH5 alpha
Culture conditions: 37 C, aerobic, LB
5'sequencing primers: M13R:CAGGAAACAGCTATGACC
3'sequencing primers: M13F:TGTAAAACGACGGCCAGT
Remarks: blue spot screening produces single strand DNA.
pUC119 Description
The pUC119 vector was constructed by inserting a HgiA I (5645) DraI (5941) fragment containing M13 phage DNA Intergenic region (IG) into the Nde I cleavage site of the pUC19 vector. Because lacZ gene contains polyclonal loci, it is easy to detect the insertion of DNA fragments in the vector by blue-and-white screening on the plate medium containing IPTG and X-Gal. At the same time, exogenous genes can be expressed by lac promoter on the vector, and DNA fragments inserted into the vector can be sequenced using M13 series of universal primers. Deletion Kit for Kilo-Sequence (Code No. 6030) can be used to sequence the DNA fragments of thousands of bases cloned.
Because of the insertion of the intergenic region (IG) of M13 phage DNA into the pUC119 vector, the pUC119 vector can also produce single stranded DNA (SSDNA) under Helper phage M13K07 infection, which is released from the bacteria after encapsulation in Phage particles without Helper phage single stranded DNA contamination. When using this product, stable single chain long fragment DNA (~7kb) can be obtained steadily, and there is no deletion.
pUC119 Mutiple cloning site
pUC119 Sequence
LOCUS Exported 3162 bp ds-DNA circular SYN 22-9-2015
KEYWORDS Untitled 4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3162)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-9-22
FEATURES Location/Qualifiers
source 1..3162
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 205..221
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 234..290
/gene="
"
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(291..307)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 520..975
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1257..1361
/gene="bla"
/note="AmpR promoter"
CDS 1362..2222
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2393..2981
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
241 catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc
301 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
361 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
421 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
481 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
541 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
601 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
661 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
721 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
781 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
841 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
901 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
961 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
1021 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
1081 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
1141 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
1201 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
1261 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
1321 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
1381 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
1441 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
1501 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
1561 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
1621 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
1681 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
1741 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
1801 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
1861 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
1921 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
1981 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
2041 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
2101 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
2161 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
2221 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
2281 ttaaaaggat ctaggtgaag atcctttttg <
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