LOCUS       pF3A_WG_(BYDV).        3689 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for cell-free 
            translation in a wheat germ extract.
ACCESSION   .
VERSION     .
KEYWORDS    pF3A WG (BYDV)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3689)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3689)
  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..3689
                     /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"
     promoter        1083..1187
                     /label=AmpR promoter
     CDS             1188..2045
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2206..2794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2910..3193
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3423..3509
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3601..3628
                     /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 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg
     1081 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1141 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1201 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1261 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1321 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1381 caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1441 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1501 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1561 taaccatgag tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg
     1621 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1681 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1741 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1801 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1861 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1921 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1981 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2041 attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     2101 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2161 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2221 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2281 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2341 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2401 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2461 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2521 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2581 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2641 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2701 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2761 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     2821 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2881 aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc
     2941 cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat
     3001 gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt
     3061 gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg
     3121 aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc
     3181 tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg
     3241 agatcttttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     3301 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     3361 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     3421 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3481 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     3541 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     3601 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     3661 taaatacatt caaatatgta tccgctcat
//
