LOCUS       pFN29K_His6HaloT        4393 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for bacterial or
            cell-free expression of a protein with a cleavable N-terminal 
            6xHis-HaloTag(R) dual tag.
ACCESSION   .
VERSION     .
KEYWORDS    pFN29K His6HaloTag T7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4393)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4393)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Subclone a coding sequence between the SgfI/AsiSI and PmeI sites to 
            remove the lethal barnase gene.
FEATURES             Location/Qualifiers
     source          1..4393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             70..72
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             76..93
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             94..981
                     /label=HaloTag(R)
                     /note="modified bacterial dehalogenase that forms covalent
                     bonds with chloroalkane derivatives"
     CDS             994..1014
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             1052..1384
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
     misc_feature    1397..1452
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      1516..1563
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1944..2735
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     rep_origin      complement(2910..3498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3614..3897
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      4127..4213
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4305..4332
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactataggg tattttaact ttactaagga
       61 gaattcatca tgaaacatca tcaccatcac cacgcagaaa tcggtactgg ctttccattc
      121 gacccccatt atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc
      181 gatggcaccc ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac
      241 atcatcccgc atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc
      301 aaatccgaca aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc
      361 ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct
      421 ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag
      481 ttcatccgcc ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag
      541 gccttccgca ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag
      601 ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc
      661 gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca
      721 atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac
      781 cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc
      841 gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt
      901 ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg
      961 tcgacgctcg agatttccgg cgagccaacc actgaggatc tgtactttca gagcgataac
     1021 gcgatcgcca tggaataagt aaggaatcca catggcacag gttatcaaca cgtttgacgg
     1081 ggttgcggat tatcttcaga catatcataa gctacctgat aattacatta caaaatcaga
     1141 agcacaagcc ctcggctggg tggcatcaaa agggaacctt gcagacgtcg ctccggggaa
     1201 aagcatcggc ggagacatct tctcaaacag ggaaggcaaa ctcccgggca aaagcggacg
     1261 aacatggcgt gaagcggata ttaactatac atcaggcttc agaaattcag accggattct
     1321 ttactcaagc gactggctga tttacaaaac aacggaccat tatcagacct ttacaaaaat
     1381 cagataatgt ttaaacgaat tcgggctcgg tacccgggga tcctctagag tcgacctgca
     1441 ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1501 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1561 ttgctgaaag gaggaactat atccggttcg cttgctgtcc ataaaaccgc ccagtctagc
     1621 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt
     1681 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     1741 ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg gcagcgtgag
     1801 cttcaaaaga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     1861 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag
     1921 agacaggatg acggtcgttt cgcatgcttg aacaagatgg attgcacgca ggttctccgg
     1981 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2041 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2101 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     2161 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2221 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2281 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2341 tcgaccacca agcgaaacat cgcatcgagc gagcacgcac tcggatggaa gccggtcttg
     2401 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2461 ggctcaaggc gcgtatgccg gatggtgagg atctcgtcgt gactcatggc gatgcctgct
     2521 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2581 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2641 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     2701 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     2761 gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc
     2821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2881 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3481 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3601 atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc tgggatggtg
     3661 aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc tgtaccctct
     3721 ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca gtttttcggg
     3781 tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt
     3841 gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag ccatcgagat
     3901 aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct tttcagcctg
     3961 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     4021 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     4081 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     4141 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     4201 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     4261 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     4321 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     4381 tgtatccgct cat
//
