LOCUS       40924_20386        4137 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for bacterial 
            or in vitro expression of a protein with a cleavable N-terminal GST 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4137)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4137)
  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..4137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             70..723
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             742..762
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQA"
     CDS             791..1123
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLQGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    1136..1191
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      1255..1302
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1531..1635
                     /label=AmpR promoter
     CDS             1636..2493
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(2654..3242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3358..3641
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3871..3957
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4049..4076
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactataggg aattttaact ttactaagga
       61 gaatttatca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
      121 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
      181 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
      241 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
      301 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
      361 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
      421 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
      481 tgtcataaaa catatttgaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
      541 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
      601 tgtttcaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
      661 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
      721 aaatccggag gtggtggcgg agaaaacctg tacttccaag cgatcgccat ggaataagta
      781 aaggaatcac atggcacagg ttatcaacac gtttgacggg gttgcggatt atcttcagac
      841 atatcataag ctacctgata attacattac aaaatcagaa gcacaagccc tcggctgggt
      901 ggcatcaaaa gggaaccttg cagacgtcgc tccggggaaa agcatcggcg gagacatctt
      961 ctcaaacagg gaaggcaaac tccagggcaa aagcggacga acatggcgtg aagcggatat
     1021 taactataca tcaggcttca gaaattcaga ccggattctt tactcaagcg actggctgat
     1081 ttacaaaaca acggaccatt atcagacctt tacaaaaatc agataatgtt taaacgaatt
     1141 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct
     1201 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     1261 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     1321 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     1381 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     1441 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtaat ggtttcttag
     1501 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1561 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1621 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1681 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1741 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1801 gagagttttc gccccgaaga acgttttcca atgatgagca ctttcaaagt tctgctatgt
     1861 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1921 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1981 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2041 cttctgacaa ctatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2101 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2161 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2221 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2281 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2341 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2401 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2461 gctgagatag gtgcctcact gattaagcat tggtaattcg aaatgaccga ccaagcgacg
     2521 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3061 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     3241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     3361 gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg
     3421 cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc
     3481 atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt
     3541 ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga
     3601 ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg
     3661 gccacgatgc gtccggcgta gaggatcgag atcttttcag cctgatacag attaaatcag
     3721 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     3781 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     3841 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     3901 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3961 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4021 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     4081 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcat
//
