LOCUS       pFN29A_His6HaloT        4398 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for bacterial 
            or cell-free expression of a protein with a cleavable N-terminal 
            6xHis-HaloTag(R) dual tag.
ACCESSION   .
VERSION     .
KEYWORDS    pFN29A His6HaloTag T7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4398)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4398)
  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..4398
                     /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"
     promoter        1792..1896
                     /label=AmpR promoter
     CDS             1897..2754
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2915..3503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3619..3902
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      4132..4218
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4310..4337
                     /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 ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1801 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1861 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1921 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1981 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2041 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2101 actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2161 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2221 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2281 gataacactg cggccaactt acttctgaca actatcggag gaccgaagga gctaaccgct
     2341 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2401 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2461 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2521 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2581 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2641 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2701 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaattc
     2761 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     2821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3421 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3481 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3541 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3601 cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga
     3661 tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac
     3721 cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt
     3781 tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta
     3841 gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc
     3901 gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatcttttca
     3961 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     4021 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     4081 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     4141 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     4201 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     4261 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     4321 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     4381 aaatatgtat ccgctcat
//
