LOCUS       40924_20391        4132 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin 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 4132)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4132)
  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..4132
                     /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"
     CDS             1683..2474
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(2649..3237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3353..3636
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3866..3952
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4044..4071
                     /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 tctacgtgtt ccgcttcctt
     1501 tagcagccct tgcgccctga gtgcttgcgg cagcgtgagc ttcaaaagaa ttgccagctg
     1561 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     1621 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga cggtcgtttc
     1681 gcatgcttga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     1741 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     1801 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     1861 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     1921 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     1981 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2041 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2101 gcatcgagcg agcacgcact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2161 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgtatgccgg
     2221 atggtgagga tctcgtcgtg actcatggcg atgcctgctt gccgaatatc atggtggaaa
     2281 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2341 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2401 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2461 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     2521 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2581 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2821 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3061 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     3241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat
     3361 acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct
     3421 tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg
     3481 gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc
     3541 tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg
     3601 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     3661 gatgcgtccg gcgtagagga tcgagatctt ttcagcctga tacagattaa atcagaacgc
     3721 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     3781 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     3841 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3901 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     3961 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     4021 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     4081 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc at
//
