pFN2K (GST)
At a Glance
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<protect>
| Type |
Cloning |
|---|---|
| Origin of Replication |
ColE1 |
| Copy # | |
| Markers |
Kanamycin |
| Link to Sequence | |
| Features |
T7 RNA polymerase promoter |
| Accession Number | |
| edit table |
</protect>
Notes
The pFN2K (GST) vector is part of a group of plasmids that are all grouped under the Flexi Vector System. This system is a powerful directional cloning methof for protein-coding sequences. The basis for the method is the two restriction enzymes SgfI(d) and PmeI(c). These are two rare-cutting restriction enzy,es that provide a rapid, efficient way to transfer protein-coding regions back and forth between the Flexi vectors. Using these restriction enzymes elimates the need to resequence. By replacing the DNA sequence of interest by means of the lethal barnase gene contained in each vector, it acts as a positive selection for he successful ligation of the insert.
Use of the Flexi Vectors eliminates the need for appending multiple amino acids to the N- or C-termini of the protein of interest like so many other vectors require. This vector system also does not require the use of an archival entry vector.
History
pFN2K (GST) is part of a group of vectors that are very similar. The only differences are the resistance gene and a few other very small differences.
Sources
References
See Help:References for how to manage references in EcoliWiki.
Sequence
1 GGATCTCGAT CCCGCGAAAT TAATACGACT CACTATAGGG AATTTTAACT 51 TTACTAAGGA GAATTTATCA TGTCCCCTAT ACTAGGTTAT TGGAAAATTA 101 AGGGCCTTGT GCAACCCACT CGACTTCTTT TGGAATATCT TGAAGAAAAA 151 TATGAAGAGC ATTTGTATGA GCGCGATGAA GGTGATAAAT GGCGAAACAA 201 AAAGTTTGAA TTGGGTTTGG AGTTTCCCAA TCTTCCTTAT TATATTGATG 251 GTGATGTTAA ATTAACACAG TCTATGGCCA TCATACGTTA TATAGCTGAC 301 AAGCACAACA TGTTGGGTGG TTGTCCAAAA GAGCGTGCAG AGATTTCAAT 351 GCTTGAAGGA GCGGTTTTGG ATATTAGATA CGGTGTTTCG AGAATTGCAT 401 ATAGTAAAGA CTTTGAAACT CTCAAAGTTG ATTTTCTTAG CAAGCTACCT 451 GAAATGCTGA AAATGTTCGA AGATCGTTTA TGTCATAAAA CATATTTGAA 501 TGGTGATCAT GTAACCCATC CTGACTTCAT GTTGTATGAC GCTCTTGATG 551 TTGTTTTATA CATGGACCCA ATGTGCCTGG ATGCGTTCCC AAAATTAGTT 601 TGTTTCAAAA AACGTATTGA AGCTATCCCA CAAATTGATA AGTACTTGAA 651 ATCCAGCAAG TATATAGCAT GGCCTTTGCA GGGCTGGCAA GCCACGTTTG 701 GTGGTGGCGA CCATCCTCCA AAATCCGGAG GTGGTGGCGG AGAAAACCTG 751 TACTTCCAAG CGATCGCCAT GGAATAAGTA AAGGAATCAC ATGGCACAGG 801 TTATCAACAC GTTTGACGGG GTTGCGGATT ATCTTCAGAC ATATCATAAG 851 CTACCTGATA ATTACATTAC AAAATCAGAA GCACAAGCCC TCGGCTGGGT 901 GGCATCAAAA GGGAACCTTG CAGACGTCGC TCCGGGGAAA AGCATCGGCG 951 GAGACATCTT CTCAAACAGG GAAGGCAAAC TCCAGGGCAA AAGCGGACGA 1001 ACATGGCGTG AAGCGGATAT TAACTATACA TCAGGCTTCA GAAATTCAGA 1051 CCGGATTCTT TACTCAAGCG ACTGGCTGAT TTACAAAACA ACGGACCATT 1101 ATCAGACCTT TACAAAAATC AGATAATGTT TAAACGAATT CGAGCTCGGT 1151 ACCCGGGGAT CCTCTAGAGT CGACCTGCAG GCATGCAAGC TGATCCGGCT 1201 GCTAACAAAG CCCGAAAGGA AGCTGAGTTG GCTGCTGCCA CCGCTGAGCA 1251 ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT 1301 TGCTGAAAGG AGGAACTATA TCCGGTTCGC TTGCTGTCCA TAAAACCGCC 1351 CAGTCTAGCT ATCGCCATGT AAGCCCACTG CAAGCTACCT GCTTTCTCTT 1401 TGCGCTTGCG TTTTCCCTTG TCCAGATAGC CCAGTAGCTG ACATTCATCC 1451 GGGGTCAGCA CCGTTTCTGC GGACTGGCTT TCTACGTGTT CCGCTTCCTT 1501 TAGCAGCCCT TGCGCCCTGA GTGCTTGCGG CAGCGTGAGC TTCAAAAGAA 1551 TTGCCAGCTG GGGCGCCCTC TGGTAAGGTT GGGAAGCCCT GCAAAGTAAA 1601 CTGGATGGCT TTCTTGCCGC CAAGGATCTG ATGGCGCAGG GGATCAAGAT 1651 CTGATCAAGA GACAGGATGA CGGTCGTTTC GCATGCTTGA ACAAGATGGA 1701 TTGCACGCAG GTTCTCCGGC CGCTTGGGTG GAGAGGCTAT TCGGCTATGA 1751 CTGGGCACAA CAGACAATCG GCTGCTCTGA TGCCGCCGTG TTCCGGCTGT 1801 CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA AGACCGACCT GTCCGGTGCC 1851 CTGAATGAAC TGCAGGACGA GGCAGCGCGG CTATCGTGGC TGGCCACGAC 1901 GGGCGTTCCT TGCGCAGCTG TGCTCGACGT TGTCACTGAA GCGGGAAGGG 1951 ACTGGCTGCT ATTGGGCGAA GTGCCGGGGC AGGATCTCCT GTCATCTCAC 2001 CTTGCTCCTG CCGAGAAAGT ATCCATCATG GCTGATGCAA TGCGGCGGCT 2051 GCATACGCTT GATCCGGCTA CCTGCCCATT CGACCACCAA GCGAAACATC 2101 GCATCGAGCG AGCACGCACT CGGATGGAAG CCGGTCTTGT CGATCAGGAT 2151 GATCTGGACG AAGAGCATCA GGGGCTCGCG CCAGCCGAAC TGTTCGCCAG 2201 GCTCAAGGCG CGTATGCCGG ATGGTGAGGA TCTCGTCGTG ACTCATGGCG 2251 ATGCCTGCTT GCCGAATATC ATGGTGGAAA ATGGCCGCTT TTCTGGATTC 2301 ATCGACTGTG GCCGGCTGGG TGTGGCGGAC CGCTATCAGG ACATAGCGTT 2351 GGCTACCCGT GATATTGCTG AAGAGCTTGG CGGCGAATGG GCTGACCGCT 2401 TCCTCGTGCT TTACGGTATC GCCGCTCCCG ATTCGCAGCG CATCGCCTTC 2451 TATCGCCTTC TTGACGAGTT CTTCTGAGCG GGACTCTGGG GTTCGAAATG 2501 ACCGACCAAG CGACGCCCAA CCGGTATCAG CTCACTCAAA GGCGGTAATA 2551 CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA 2601 AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT 2651 TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT 2701 CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC 2751 TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT 2801 ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCATAGCTCA 2851 CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG 2901 TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT 2951 ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA 3001 GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA 3051 GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGG ACAGTATTTG 3101 GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC 3151 TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG 3201 CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATTTCAAGAA GATCCTTTGA 3251 TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG 3301 ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTATA 3351 GTCCGGAAAT ACAGGAACGC ACGCTGGATG GCCCTTCGCT GGGATGGTGA 3401 AACCATGAAA AATGGCAGCT TCAGTGGATT AAGTGGGGGT AATGTGGCCT 3451 GTACCCTCTG GTTGCATAGG TATTCATACG GTTAAAATTT ATCAGGCGCG 3501 ATTGCGGCAG TTTTTCGGGT GGTTTGTTGC CATTTTTACC TGTCTGCTGC 3551 CGTGATCGCG CTGAACGCGT TTTAGCGGTG CGTACAATTA AGGGATTATG 3601 GTAAATCCAC TTACTGTCTG CCCTCGTAGC CATCGAGATA AACCGCAGTA 3651 CTCCGGCCAC GATGCGTCCG GCGTAGAGGA TCGAGATCTT TTCAGCCTGA 3701 TACAGATTAA ATCAGAACGC AGAAGCGGTC TGATAAAACA GAATTTGCCT 3751 GGCGGCAGTA GCGCGGTGGT CCCACCTGAC CCCATGCCGA ACTCAGAAGT 3801 GAAACGCCGT AGCGCCGATG GTAGTGTGGG GTCTCCCCAT GCGAGAGTAG 3851 GGAACTGCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC 3901 CTTTCGTTTT ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA 3951 ATCCGCCGGG AGCGGATTTG AACGTTGCGA AGCAACGGCC CGGAGGGTGG 4001 CGGGCAGGAC GCCCGCCATA AACTGCCAGG CATCAAATTA AGCAGAAGGC 4051 CATCCTGACG GATGGCCTTT TTGCGTTTCT ACAAACTCTT TTGTTTATTT 4101 TTCTAAATAC ATTCAAATAT GTATCCGCTC AT
