LOCUS       40924_20341        4128 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for mammalian 
            expression from a strongly compromised CMV promoter of a protein 
            with an N-terminal HaloTag(R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4128)
  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..4128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..45
                     /label=CMVd3 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields strongly reduced expression 
                     relative to the full-length promoter"
     promoter        77..95
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        101..119
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             136..1026
                     /codon_start=1
                     /label=HaloTag(R)
                     /note="modified bacterial dehalogenase that forms covalent
                     bonds with chloroalkane derivatives"
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1039..1059
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="EDLYFQS"
     CDS             1097..1429
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    1442..1497
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     polyA_signal    complement(1602..1723)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2122..2913
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(3088..3676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3792..4075
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
       61 cagatcacta gaagcttaat acgactcact atagggtacg atttaggtga cactatagaa
      121 taaggagaat tcatcatggc agaaatcggt actggctttc cattcgaccc ccattatgtg
      181 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
      241 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
      301 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
      361 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
      421 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
      481 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
      541 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
      601 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
      661 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
      721 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
      781 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
      841 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
      901 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
      961 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt
     1021 tccggcgagc caaccactga ggatctgtac tttcagagcg ataacgcgat cgccatggaa
     1081 taagtaagga atccacatgg cacaggttat caacacgttt gacggggttg cggattatct
     1141 tcagacatat cataagctac ctgataatta cattacaaaa tcagaagcac aagccctcgg
     1201 ctgggtggca tcaaaaggga accttgcaga cgtcgctccg gggaaaagca tcggcggaga
     1261 catcttctca aacagggaag gcaaactccc gggcaaaagc ggacgaacat ggcgtgaagc
     1321 ggatattaac tatacatcag gcttcagaaa ttcagaccgg attctttact caagcgactg
     1381 gctgatttac aaaacaacgg accattatca gacctttaca aaaatcagat aatgtttaaa
     1441 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     1501 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1561 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     1621 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1681 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1741 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     1801 ctacaaatgt ggtaaaatcg aattctaatg gatcctcttt gcgcttgcgt tttcccttgt
     1861 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     1921 ctacgtgttc cgcttccttt agcagccctt gcgccctgag tgcttgcggc agcgtgagct
     1981 tcaaaagaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     2041 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag
     2101 acaggatgac ggtcgtttcg catgcttgaa caagatggat tgcacgcagg ttctccggcc
     2161 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2221 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2281 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     2341 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2401 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2461 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2521 gaccaccaag cgaaacatcg catcgagcga gcacgcactc ggatggaagc cggtcttgtc
     2581 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     2641 ctcaaggcgc gtatgccgga tggtgaggat ctcgtcgtga ctcatggcga tgcctgcttg
     2701 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     2761 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     2821 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     2881 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     2941 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3661 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3781 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     3841 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     3901 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     3961 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     4021 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     4081 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
