LOCUS       pF3K_WG_(BYDV).        3684 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for cell-free 
            translation in a wheat germ extract.
ACCESSION   .
VERSION     .
KEYWORDS    pF3K WG (BYDV)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3684)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3684)
  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..3684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..41
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        46..63
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           65..201
                     /label=BYDV 5' UTR
                     /note="barley yellow dwarf virus 5' untranslated region; 
                     required for function of the cap-independent 3' translation
                     enhancer (Guo et al., 2001)"
     CDS             233..565
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
     misc_feature    596..702
                     /label=BYDV 3' TE
                     /note="cap-independent 3' translation enhancer from barley 
                     yellow dwarf virus (Wang et al., 1997)"
     terminator      807..854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1235..2026
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     rep_origin      complement(2201..2789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2905..3188
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3418..3504
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3596..3623
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatttaggt gacactatag aagggtaata cgactcacta
       61 tagcagtgaa gattgaccat ctcacaaaag ctgttacgtg cttgtaacac actacacact
      121 cgttttgtat tcgagaagta gttgcaacaa cggtcccctt attgcctgac aagctgaggg
      181 ccacccttct atccccaccg cgcgatcgcc atggaataag taaggaatcc acatggcaca
      241 ggttatcaac acgtttgacg gggttgcgga ttatcttcag acatatcata agctacctga
      301 taattacatt acaaaatcag aagcacaagc cctcggctgg gtggcatcaa aagggaacct
      361 tgcagacgtc gctccgggga aaagcatcgg cggagacatc ttctcaaaca gggaaggcaa
      421 actccagggc aaaagcggac gaacatggcg tgaagcggat attaactata catcaggctt
      481 cagaaattca gaccggattc tttactcaag cgactggctg atttacaaaa caacggacca
      541 ttatcagacc tttacaaaaa tcagataatg tttaaacgaa ttcgagctcg gtaccagtga
      601 agacaacacc actagcacaa atcggatcct gggaaacagg cagaacttcg gttcataagc
      661 tcgggtaggc tgtcaaccta ccgccgtatc gtattgtgtt tgtctagata cgtatcgcga
      721 gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
      781 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      841 tgaggggttt tttgctgaaa ggaggaacta tatccggttc gcttgctgtc cataaaaccg
      901 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
      961 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     1021 gcggactggc tttctacgtg ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc
     1081 ggcagcgtga gcttcaaaag aattgccagc tggggcgccc tctggtaagg ttgggaagcc
     1141 ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag
     1201 atctgatcaa gagacaggat gacggtcgtt tcgcatgctt gaacaagatg gattgcacgc
     1261 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     1321 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     1381 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     1441 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     1501 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     1561 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     1621 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgca ctcggatgga
     1681 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     1741 actgttcgcc aggctcaagg cgcgtatgcc ggatggtgag gatctcgtcg tgactcatgg
     1801 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     1861 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     1921 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     1981 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     2041 gggttcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     2101 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2161 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2221 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2281 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2341 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2401 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2461 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2521 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2581 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2641 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2701 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2761 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     2821 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2881 tcttcaccta gatcctttta tagtccggaa atacaggaac gcacgctgga tggcccttcg
     2941 ctgggatggt gaaaccatga aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc
     3001 ctgtaccctc tggttgcata ggtattcata cggttaaaat ttatcaggcg cgattgcggc
     3061 agtttttcgg gtggtttgtt gccattttta cctgtctgct gccgtgatcg cgctgaacgc
     3121 gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta
     3181 gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc
     3241 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     3301 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     3361 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     3421 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3481 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     3541 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     3601 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     3661 acattcaaat atgtatccgc tcat
//
