pET-30 Xa/LIC
At a Glance
Image of pET-30 Xa/LIC. Novagen
<protect>
| Type |
Cloning |
|---|---|
| Origin of Replication |
f1 |
| Copy # |
~40 |
| Markers |
Kanamycin |
| 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.
History
The pET-30 Xa/LIC vector is an Xa/LIC version of pET-30b(+).
Sources
References
See Help:References for how to manage references in EcoliWiki.
- ↑ 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
- ↑ 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 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
