LOCUS       pF25K_ICE_T7.        3942 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for cell-free 
            translation in an insect cell extract.
ACCESSION   .
VERSION     .
KEYWORDS    pF25K ICE T7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3942)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3942)
  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..3942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           40..85
                     /label=polyhedrin 5' UTR
                     /note="5' UTR from the Malacosoma neustria
                     nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)"
     CDS             117..449
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
     misc_feature    462..517
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     3'UTR           611..983
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     misc_feature    988..1017
                     /label=poly(A)
                     /note="poly(A)"
                     /note="synthetic poly(A) tract"
     terminator      1018..1065
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1493..2284
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     rep_origin      complement(2459..3047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3163..3446
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3676..3762
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3854..3881
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactatagga gtatttttat tctttcgtaa
       61 aaaaattaga aaaataaaat ataaagcgat cgccatggaa taagtaagga atccacatgg
      121 cacaggttat caacacgttt gacggggttg cggattatct tcagacatat cataagctac
      181 ctgataatta cattacaaaa tcagaagcac aagccctcgg ctgggtggca tcaaaaggga
      241 accttgcaga cgtcgctccg gggaaaagca tcggcggaga catcttctca aacagggaag
      301 gcaaactccc gggcaaaagc ggacgaacat ggcgtgaagc ggatattaac tatacatcag
      361 gcttcagaaa ttcagaccgg attctttact caagcgactg gctgatttac aaaacaacgg
      421 accattatca gacctttaca aaaatcagat aatgtttaaa cgaattcggg ctcggtaccc
      481 ggggatcctc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
      541 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac ctagagtcgg
      601 gcggctgtaa aacacgatac attgttatta gtacatttat taagcgctag attctgtgcg
      661 ttgttgattt acagacaatt gttgtacgta ttttaataat tcattaaatt tataatcttt
      721 agggtggtat gttagagcga aaatcaaatg attttcagcg tctttatatc tgaatttaaa
      781 tattaaatcc tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca agggttgttt
      841 ttccgaaccg atggctggac tatctaatgg attttcgctc aacgccacaa aacttgccaa
      901 atcttgtagc agcaatctag ctttgtcgat attcgtttgt gttttgtttt gtaataaagg
      961 ttcgacgtcg ttcaaaatat tatccgcaaa aaaaaaaaaa aaaaaaaaaa aaaaaaacta
     1021 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctcgc gagcggccgc
     1081 aggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggttcgc
     1141 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     1201 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     1261 ggggtcagca ccgtttctgc ggactggctt tctacgtgtt ccgcttcctt tagcagccct
     1321 tgcgccctga gtgcttgcgg cagcgtgagc ttcaaaagaa ttgccagctg gggcgccctc
     1381 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     1441 atggcgcagg ggatcaagat ctgatcaaga gacaggatga cggtcgtttc gcatgcttga
     1501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     1921 agcacgcact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgtatgccgg atggtgagga
     2041 tctcgtcgtg actcatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2281 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa ccggtatcag
     2341 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2401 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2461 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2521 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2581 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2641 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2701 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2761 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2821 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2881 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2941 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3001 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga
     3061 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3121 tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat acaggaacgc
     3181 acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct tcagtggatt
     3241 aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg gttaaaattt
     3301 atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc tgtctgctgc
     3361 cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg gtaaatccac
     3421 ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac gatgcgtccg
     3481 gcgtagagga tcgagatctt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     3541 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     3601 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     3661 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     3721 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     3781 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     3841 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     3901 ttgtttattt ttctaaatac attcaaatat gtatccgctc at
//
