pF1A T7

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At a Glance

pF1A T7, Image made with PlasMapper

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Type

Cloning
Flexi(R) Vector

Origin of Replication

ColE1

Copy #
Markers

Ampicillin

Link to Sequence

NCBI

Features

T7 Promoter
Sgf I site
Barnase coding region
Pme I site
T7 Terminator
rrnB transcription terminator

Accession Number

AY753576

edit table

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Notes

The pF1A T7 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

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Sources

Promega

References

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Sequence

  
    1  GGATCTCGAT CCCGCGAAAT TAATACGACT CACTATAGGG GAATTGTGAG
   51  CGGATAACAA TTCCCCACTA GTAATAATTT TCTTTAACTT TAGTAAGGAG
  101  CGATCGCCAT GGCACAGGTT ATCAACACGT TTGACGGGGT TGCGGATTAT
  151  CTTCAGACAT ATCATAAGCT ACCTGATAAT TACATTACAA AATCAGAAGC
  201  ACAAGCCCTC GGCTGGGTGG CATCAAAAGG GAACCTTGCA GACGTCGCTC
  251  CGGGGAAAAG CATCGGCGGA GACATCTTCT CAAACAGGGA AGGCAAACTC
  301  CAGGGCAAAA GCGGACGAAC ATGGCGTGAA GCGGATATTA ACTATACATC
  351  AGGCTTCAGA AATTCAGACC GGATTCTTTA CTCAAGCGAC TGGCTGATTT  
  401  ACAAAACAAC GGACCATTAT CAGACCTTTA CAAAAATCAG ATAATGTTTA
  451  AACGAATTCG AGCTCGGTAC CCGGGGATCC TCTAGAGTCG ACCTGCAGGC
  501  ATGCAAGCTG ATCCGGCTGC TAACAAAGCC CGAAAGGAAG CTGAGTTGGC
  551  TGCTGCCACC GCTGAGCAAT AACTAGCATA ACCCCTTGGG GCCTCTAAAC
  601  GGGTCTTGAG GGGTTTTTTG CTGAAAGGAG GAACTATATC CGGTTCGCTT
  651  GCTGTCCATA AAACCGCCCA GTCTAGCTAT CGCCATGTAA GCCCACTGCA
  701  AGCTACCTGC TTTCTCTTTG CGCTTGCGTT TTCCCTTGTC CAGATAGCCC
  751  AGTAGCTGAC ATTCATCCGG GGTCAGCACC GTTTCTGCGG ACTGGCTTTC
  801  TACGTAATGG TTTCTTAGAC GTCAGGTGGC ACTTTTCGGG GAAATGTGCG
  851  CGGAACCCCT ATTTGTTTAT TTTTCTAAAT ACATTCAAAT ATGTATCCGC
  901  TCATGAGACA ATAACCCTGA TAAATGCTTC AATAATATTG AAAAAGGAAG
  951  AGTATGAGTA TTCAACATTT CCGTGTCGCC CTTATTCCCT TTTTTGCGGC
 1001  ATTTTGCCTT CCTGTTTTTG CTCACCCAGA AACGCTGGTG AAAGTAAAAG
 1051  ATGCTGAAGA TCAGTTGGGT GCACGAGTGG GTTACATCGA ACTGGATCTC
 1101  AACAGCGGTA AGATCCTTGA GAGTTTTCGC CCCGAAGAAC GTTTTCCAAT
 1151  GATGAGCACT TTCAAAGTTC TGCTATGTGG CGCGGTATTA TCCCGTATTG
 1201  ACGCCGGGCA AGAGCAACTC GGTCGCCGCA TACACTATTC TCAGAATGAC
 1251  TTGGTTGAGT ACTCACCAGT CACAGAAAAG CATCTTACGG ATGGCATGAC
 1301  AGTAAGAGAA TTATGCAGTG CTGCCATAAC CATGAGTGAT AACACTGCGG
 1351  CCAACTTACT TCTGACAACT ATCGGAGGAC CGAAGGAGCT AACCGCTTTT
 1401  TTGCACAACA TGGGGGATCA TGTAACTCGC CTTGATCGTT GGGAACCGGA
 1451  GCTGAATGAA GCCATACCAA ACGACGAGCG TGACACCACG ATGCCTGTAG
 1501  CAATGGCAAC AACGTTGCGC AAACTATTAA CTGGCGAACT ACTTACTCTA
 1551  GCTTCCCGGC AACAATTAAT AGACTGGATG GAGGCGGATA AAGTTGCAGG
 1601  ACCACTTCTG CGCTCGGCCC TTCCGGCTGG CTGGTTTATT GCTGATAAAT
 1651  CTGGAGCCGG TGAGCGTGGG TCTCGCGGTA TCATTGCAGC ACTGGGGCCA
 1701  GATGGTAAGC CCTCCCGTAT CGTAGTTATC TACACGACGG GGAGTCAGGC
 1751  AACTATGGAT GAACGAAATA GACAGATCGC TGAGATAGGT GCCTCACTGA
 1801  TTAAGCATTG GTAATTCGAA ATGACCGACC AAGCGACGCC CAACCGGTAT
 1851  CAGCTCACTC AAAGGCGGTA ATACGGTTAT CCACAGAATC AGGGGATAAC
 1901  GCAGGAAAGA ACATGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA
 1951  AAAGGCCGCG TTGCTGGCGT TTTTCCATAG GCTCCGCCCC CCTGACGAGC
 2001  ATCACAAAAA TCGACGCTCA AGTCAGAGGT GGCGAAACCC GACAGGACTA
 2051  TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC GCTCTCCTGT
 2101  TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA
 2151  GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG
 2201  GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA
 2251  CCGCTGCGCC TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGTAAGAC
 2301  ACGACTTATC GCCACTGGCA GCAGCCACTG GTAACAGGAT TAGCAGAGCG
 2351  AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC CTAACTACGG
 2401  CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA
 2451  CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT
 2501  GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA
 2551  AGGATTTCAA GAAGATCCTT TGATCTTTTC TACGGGGTCT GACGCTCAGT
 2601  GGAACGAAAA CTCACGTTAA GGGATTTTGG TCATGAGATT ATCAAAAAGG
 2651  ATCTTCACCT AGATCCTTTT ATAGTCCGGA AATACAGGAA CGCACGCTGG
 2701  ATGGCCCTTC GCTGGGATGG TGAAACCATG AAAAATGGCA GCTTCAGTGG
 2751  ATTAAGTGGG GGTAATGTGG CCTGTACCCT CTGGTTGCAT AGGTATTCAT
 2801  ACGGTTAAAA TTTATCAGGC GCGATTGCGG CAGTTTTTCG GGTGGTTTGT
 2851  TGCCATTTTT ACCTGTCTGC TGCCGTGATC GCGCTGAACG CGTTTTAGCG
 2901  GTGCGTACAA TTAAGGGATT ATGGTAAATC CACTTACTGT CTGCCCTCGT
 2951  AGCCATCGAG ATAAACCGCA GTACTCCGGC CACGATGCGT CCGGCGTAGA
 3001  GGATCGAGAT CTTTTCAGCC TGATACAGAT TAAATCAGAA CGCAGAAGCG
 3051  GTCTGATAAA ACAGAATTTG CCTGGCGGCA GTAGCGCGGT GGTCCCACCT
 3101  GACCCCATGC CGAACTCAGA AGTGAAACGC CGTAGCGCCG ATGGTAGTGT
 3151  GGGGTCTCCC CATGCGAGAG TAGGGAACTG CCAGGCATCA AATAAAACGA
 3201  AAGGCTCAGT CGAAAGACTG GGCCTTTCGT TTTATCTGTT GTTTGTCGGT
 3251  GAACGCTCTC CTGAGTAGGA CAAATCCGCC GGGAGCGGAT TTGAACGTTG
 3301  CGAAGCAACG GCCCGGAGGG TGGCGGGCAG GACGCCCGCC ATAAACTGCC
 3351  AGGCATCAAA TTAAGCAGAA GGCCATCCTG ACGGATGGCC TTTTTGCGTT
 3401  TCTACAAACT CTTTTGTTTA TTTTTCTAAA TACATTCAAA TATGTATCCG
 3451  CTCAT
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