LOCUS       40924_20316        4188 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for mammalian 
            expression from a slightly 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 4188)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4188)
  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..4188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..100
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     promoter        132..150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        156..174
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             191..1081
                     /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             1094..1114
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="EDLYFQS"
     CDS             1152..1484
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    1497..1552
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     polyA_signal    complement(1657..1778)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2025..2129
                     /label=AmpR promoter
     CDS             2130..2987
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3148..3736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3852..4135
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
       61 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      121 cactagaagc ttaatacgac tcactatagg gtacgattta ggtgacacta tagaataagg
      181 agaattcatc atggcagaaa tcggtactgg ctttccattc gacccccatt atgtggaagt
      241 cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt
      301 cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc
      361 gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct
      421 gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct
      481 ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa
      541 gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac
      601 ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt
      661 cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt
      721 cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt
      781 tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa
      841 catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct
      901 gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa
      961 aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga
     1021 caacccggac ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg agatttccgg
     1081 cgagccaacc actgaggatc tgtactttca gagcgataac gcgatcgcca tggaataagt
     1141 aaggaatcca catggcacag gttatcaaca cgtttgacgg ggttgcggat tatcttcaga
     1201 catatcataa gctacctgat aattacatta caaaatcaga agcacaagcc ctcggctggg
     1261 tggcatcaaa agggaacctt gcagacgtcg ctccggggaa aagcatcggc ggagacatct
     1321 tctcaaacag ggaaggcaaa ctcccgggca aaagcggacg aacatggcgt gaagcggata
     1381 ttaactatac atcaggcttc agaaattcag accggattct ttactcaagc gactggctga
     1441 tttacaaaac aacggaccat tatcagacct ttacaaaaat cagataatgt ttaaacgaat
     1501 tcgggctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag ctgatccggc
     1561 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1621 ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     1681 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1741 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1801 ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca
     1861 aatgtggtaa aatcgaattc taatggatcc tctttgcgct tgcgttttcc cttgtccaga
     1921 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     1981 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2041 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2101 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2161 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2221 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2281 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     2341 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2401 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2461 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2521 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     2581 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2641 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2701 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2761 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2821 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2881 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2941 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     3001 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3061 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3721 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3841 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     3901 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     3961 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     4021 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     4081 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     4141 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
