LOCUS       pF25A_ICE_T7.        3947 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for cell-free 
            translation in an insect cell extract.
ACCESSION   .
VERSION     .
KEYWORDS    pF25A ICE T7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3947)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3947)
  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..3947
                     /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"
     promoter        1341..1445
                     /label=AmpR promoter
     CDS             1446..2303
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2464..3052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3168..3451
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3681..3767
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3859..3886
                     /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 tctacgtaat ggtttcttag acgtcaggtg
     1321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1381 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1441 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1501 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1561 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1621 gccccgaaga acgttttcca atgatgagca ctttcaaagt tctgctatgt ggcgcggtat
     1681 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1741 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1801 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1861 ctatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1921 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1981 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2041 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2101 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2161 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2221 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2281 gtgcctcact gattaagcat tggtaattcg aaatgaccga ccaagcgacg cccaaccggt
     2341 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2401 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2881 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     3061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     3181 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     3241 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     3301 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     3361 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     3421 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     3481 gtccggcgta gaggatcgag atcttttcag cctgatacag attaaatcag aacgcagaag
     3541 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     3601 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     3661 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3721 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     3781 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     3841 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     3901 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcat
//
