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pET-30 Xa/LIC

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At a Glance

Image of pET-30 Xa/LIC. Novagen

Type

Cloning

Origin of Replication

f1

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 promoter
T7 transcription start
His•Tag[1]
S•Tag[1]
Multiple cloning sites
T7 terminator
lacI coding sequence
pBR322 origin

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.

History

The pET-30 Xa/LIC vector is an Xa/LIC version of pET-30b(+).

Sources

Novagen

References

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

  1. 1.0 1.1 Stols, L et al. (2002) A new vector for high-throughput, ligation-independent cloning encoding a tobacco etch virus protease cleavage site. Protein Expr. Purif. 25 8-15 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 catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga 
     421 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc 
     481 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt 
     541 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct 
     601 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg 
     661 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact 
     721 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca 
     781 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca 
     841 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt 
     901 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa 
     961 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg 
    1021 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt 
    1081 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt 
    1141 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc 
    1201 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg 
    1261 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca 
    1321 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat 
    1381 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc 
    1441 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg 
    1501 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca 
    1561 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca 
    1621 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg 
    1681 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg 
    1741 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc 
    1801 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg 
    1861 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 
    1921 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga 
    1981 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc 
    2041 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg 
    2101 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc 
    2161 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt 
    2221 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg 
    2281 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact 
    2341 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg 
    2401 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa 
    2461 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg 
    2521 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt 
    2581 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac 
    2641 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc 
    2701 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa 
    2761 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc 
    2821 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat 
    2881 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 
    2941 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt 
    3001 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc 
    3061 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 
    3121 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 
    3181 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 
    3241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 
    3301 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 
    3361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 
    3421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 
    3481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 
    3541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 
    3601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 
    3661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 
    3721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 
    3781 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 
    3841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 
    3901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 
    3961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 
    4021 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc 
    4081 atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt 
    4141 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat 
    4201 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca 
    4261 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga 
    4321 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg 
    4381 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg 
    4441 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca 
    4501 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg 
    4561 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc 
    4621 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata 
    4681 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg 
    4741 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat 
    4801 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt 
    4861 ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa 
    4921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 
    4981 gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca 
    5041 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag 
    5101 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc 
    5161 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc 
    5221 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc 
    5281 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa 
    5341 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc 
    5401 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca