LOCUS       40924_19806        4129 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for mammalian 
            expression from a moderately compromised CMV promoter of a protein 
            with a cleavable C-terminal HaloTag(R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4129)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4129)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Subclone a coding sequence between the SgfI/AsiSI and 
            EcoICRI/Eco53kI sites to remove the lethal barnase gene.
FEATURES             Location/Qualifiers
     source          1..4129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..52
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     promoter        84..102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        108..126
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             165..497
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     CDS             540..560
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="EDLYFQS"
     CDS             573..1460
                     /codon_start=1
                     /label=HaloTag(R)
                     /note="modified bacterial dehalogenase that forms covalent
                     bonds with chloroalkane derivatives"
                     /translation="SEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     polyA_signal    complement(1603..1724)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2123..2914
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(3089..3677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3793..4076
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
       61 gaaccgtcag atcactagaa gcttaatacg actcactata gggtacgatt taggtgacac
      121 tatagaataa ggagcgatcg ccatggaata agtaaggaat ccacatggca caggttatca
      181 acacgtttga cggggttgcg gattatcttc agacatatca taagctacct gataattaca
      241 ttacaaaatc agaagcacaa gccctcggct gggtggcatc aaaagggaac cttgcagacg
      301 tcgctccggg gaaaagcatc ggcggagaca tcttctcaaa cagggaaggc aaactcccgg
      361 gcaaaagcgg acgaacatgg cgtgaagcgg atattaacta tacatcaggc ttcagaaatt
      421 cagaccggat tctttactca agcgactggc tgatttacaa aacaacggac cattatcaga
      481 cctttacaaa aatcagataa tgtttaatga ccccgtgtcg agctctcgag ccaaccactg
      541 aggatctgta ctttcagagc gataacgatg gatccgaaat cggtactggc tttccattcg
      601 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
      661 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
      721 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
      781 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
      841 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
      901 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
      961 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1021 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1081 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1141 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1201 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1261 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1321 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1381 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     1441 ctactctgga gatttccggt taatagaatt ctagagtcga cctgcaggca tgcaagctga
     1501 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1561 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     1621 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1681 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1741 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     1801 tctacaaatg tggtaaaatc gaattctaat ggatcctctt tgcgcttgcg ttttcccttg
     1861 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     1921 tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg cagcgtgagc
     1981 ttcaaaagaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     2041 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
     2101 gacaggatga cggtcgtttc gcatgcttga acaagatgga ttgcacgcag gttctccggc
     2161 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2221 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2281 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     2341 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2401 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2461 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2521 cgaccaccaa gcgaaacatc gcatcgagcg agcacgcact cggatggaag ccggtcttgt
     2581 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2641 gctcaaggcg cgtatgccgg atggtgagga tctcgtcgtg actcatggcg atgcctgctt
     2701 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2761 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     2821 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     2881 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     2941 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3661 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3781 tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga
     3841 aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg
     3901 gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt
     3961 ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg
     4021 cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata
     4081 aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
