LOCUS       40924_19866        3420 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for bacterial or
            in vitro expression of a protein with a C-terminal HQ metal affinity
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3420)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3420)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Subclone a coding sequence between the SgfI/AsiSI and 
            EcoICRI/Eco53kI sites to remove the lethal barnase gene.
FEATURES             Location/Qualifiers
     source          1..3420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             70..402
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     CDS             430..447
                     /codon_start=1
                     /label=HQ tag
                     /note="HQHQHQ metal affinity tag"
                     /translation="HQHQHQ"
     terminator      543..590
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             971..1762
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(1937..2525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2641..2924
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3154..3240
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3332..3359
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactatagga aattttaact ttactaagga
       61 gcgatcgcca tggcacaggt tatcaacacg tttgacgggg ttgcggatta tcttcagaca
      121 tatcataagc tacctgataa ttacattaca aaatcagaag cacaagccct cggctgggtg
      181 gcatcaaaag ggaaccttgc agacgtcgct ccggggaaaa gcatcggcgg agacatcttc
      241 tcaaacaggg aaggcaaact cccgggcaaa agcggacgaa catggcgtga agcggatatt
      301 aactatacat caggcttcag aaattcagac cggattcttt actcaagcga ctggctgatt
      361 tacaaaacaa cggaccatta tcagaccttt acaaaaatca gataatgttt aatgaccccg
      421 tgtcgagctc atcaacacca acatcaataa tctagagtcg acctgcaggc atgcaagctg
      481 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      541 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      601 gaactatatc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
      661 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
      721 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc
      781 gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgagctt caaaagaatt
      841 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt
      901 cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgacg
      961 gtcgtttcgc atgcttgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     1021 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     1081 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     1141 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     1201 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     1261 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     1321 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     1381 gaaacatcgc atcgagcgag cacgcactcg gatggaagcc ggtcttgtcg atcaggatga
     1441 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     1501 tatgccggat ggtgaggatc tcgtcgtgac tcatggcgat gcctgcttgc cgaatatcat
     1561 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     1621 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     1681 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     1741 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     1801 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1861 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1921 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1981 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2041 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2101 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2161 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2221 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2281 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2341 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2401 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2461 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2521 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2581 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     2641 ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa
     2701 tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta
     2761 ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca
     2821 tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag
     2881 ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact
     2941 ccggccacga tgcgtccggc gtagaggatc gagatctttt cagcctgata cagattaaat
     3001 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     3061 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     3121 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3181 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3241 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     3301 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     3361 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
//
