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pET-32 Xa/LIC

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At a Glance

Image of pET-32 Xa/LIC. Novagen

Type

Cloning

Origin of Replication

f1

Copy #

~40

Markers

Ampicillin

Link to Sequence

Novagen

Features

T7 promoter
T7 transcription start
Trx•Tag[1]
His•Tag[1]
S•Tag[1]
Multiple cloning sites
lacI coding sequence

Accession Number




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

Novagen

References

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See Help:References for how to manage references in EcoliWiki.

  1. 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 EcoliWiki page
  2. Aslanidis, C & de Jong, PJ (1990) Ligation-independent cloning of PCR products (LIC-PCR). Nucleic Acids Res. 18 6069-74 PubMed EcoliWiki page
  3. Haun, RS & Moss, J (1992) Ligation-independent cloning of glutathione S-transferase fusion genes for expression in Escherichia coli. Gene 112 37-43 PubMed EcoliWiki page

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