LOCUS       40924_19041        3450 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for bacterial or
            in vitro expression of an untagged protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3450)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3450)
  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..3450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             109..441
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLQGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    454..509
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      573..620
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1001..1792
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(1967..2555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2671..2954
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3184..3270
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3362..3389
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttccccacta gtaataattt tctttaactt tagtaaggag cgatcgccat ggcacaggtt
      121 atcaacacgt ttgacggggt tgcggattat cttcagacat atcataagct acctgataat
      181 tacattacaa aatcagaagc acaagccctc ggctgggtgg catcaaaagg gaaccttgca
      241 gacgtcgctc cggggaaaag catcggcgga gacatcttct caaacaggga aggcaaactc
      301 cagggcaaaa gcggacgaac atggcgtgaa gcggatatta actatacatc aggcttcaga
      361 aattcagacc ggattcttta ctcaagcgac tggctgattt acaaaacaac ggaccattat
      421 cagaccttta caaaaatcag ataatgttta aacgaattcg agctcggtac ccggggatcc
      481 tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
      541 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      601 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggttcgctt gctgtccata
      661 aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg
      721 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
      781 gtttctgcgg actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt
      841 gcttgcggca gcgtgagctt caaaagaatt gccagctggg gcgccctctg gtaaggttgg
      901 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
      961 atcaagatct gatcaagaga caggatgacg gtcgtttcgc atgcttgaac aagatggatt
     1021 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1081 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1141 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1201 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1261 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1321 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1381 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgcactcg
     1441 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1501 agccgaactg ttcgccaggc tcaaggcgcg tatgccggat ggtgaggatc tcgtcgtgac
     1561 tcatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1621 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1681 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1741 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1801 actctggggt tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     1861 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2401 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2521 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2641 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     2701 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     2761 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     2821 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     2881 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     2941 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     3001 gagatctttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     3061 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     3121 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     3181 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3241 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     3301 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3361 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3421 ctaaatacat tcaaatatgt atccgctcat
//
