pET-32 Xa/LIC
At a Glance
Image of pET-32 Xa/LIC. Novagen
<protect>
| Type |
Cloning |
|---|---|
| Origin of Replication |
f1 |
| Copy # |
~40 |
| Markers |
Ampicillin |
| Link to Sequence | |
| Features |
T7 promoter |
| Accession Number |
|
| edit table |
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Notes
Ligation-independent cloning (LIC) was developed for directional cloning of PCR products without restriction enzyme digestion or ligation reactions[2][3] The LIC method takes advantage of the 3′ → 5′ exonuclease activity of T4 DNA polymerase to create very specific 12 to 15 nucleotide single stranded overhangs in the vector and the insert. The Srf I and Sma I sites are destroyed during Ligation Independent Cloning.
History
The pBAC-2cp Ek/LIC vector is an Ek/LIC version of pBAC-2cp.
Sources
References
See Help:References for how to manage references in EcoliWiki.
- ↑ 1.0 1.1 1.2 Eide, EJ et al. (2005) Casein kinase I in the mammalian circadian clock. Meth. Enzymol. 393 408-18 PubMed
- ↑ Aslanidis, C & de Jong, PJ (1990) Ligation-independent cloning of PCR products (LIC-PCR). Nucleic Acids Res. 18 6069-74 PubMed
- ↑ Haun, RS & Moss, J (1992) Ligation-independent cloning of glutathione S-transferase fusion genes for expression in Escherichia coli. Gene 112 37-43 PubMed
Sequence
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgatatcg ccatggccag aggagagtta gagcgaccct
241 caataccgga gccaccaccg gtacccagat ctgggctgtc catgtgctgg cgttcgaatt
301 tagcagcagc ggtttctttc ataccagaac cgcgtggcac cagaccagaa gaatgatgat
361 gatgatggtg catatggcca gaaccagaac cggccaggtt agcgtcgagg aactctttca
421 actgaccttt agacagtgca cccactttgg ttgccgccac ttcaccgttt ttgaacagca
481 gcagagtcgg gataccacgg atgccatatt tcggcgcagt gccagggttt tgatcgatgt
541 tcagttttgc aacggtcagt ttgccctgat attcgtcagc gatttcatcc agaatcgggg
601 cgatcatttt gcacggaccg caccactctg cccagaaatc gacgaggatc gccccgtccg
661 ctttgagtac atccgtgtca aaactgtcgt cagtcaggtg aataatttta tcgctcatat
721 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
781 caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta
841 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
901 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
961 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
1021 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
1081 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
1141 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
1201 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
1261 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
1321 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
1381 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
1441 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
1501 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
1561 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
1621 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
1681 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
1741 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
1801 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
1861 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
1921 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
1981 tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
2041 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
2101 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
2161 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
2221 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
2281 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
2341 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
2401 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
2461 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
2521 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
2581 ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg
2641 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
2701 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
2761 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg
2821 ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca
2881 cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc
2941 cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca
3001 tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac
3061 agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg
3121 gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg
3181 gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc
3241 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
3301 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
3361 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
3421 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg
3481 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
3541 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
3601 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
3661 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
3721 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
3781 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
3841 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
3901 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
3961 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
4021 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
4081 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
4141 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
4201 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
4261 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
4321 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
4381 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
4441 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
4501 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
4561 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
4621 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
4681 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
4741 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc
4801 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
4861 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
4921 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
4981 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
5041 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
5101 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
5161 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
5221 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
5281 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
5341 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
5461 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
5521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
5581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
5641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
5701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
5761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
5821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
5881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
