LOCUS       40924_20336        4133 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin 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 4133)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4133)
  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..4133
                     /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"
     promoter        1970..2074
                     /label=AmpR promoter
     CDS             2075..2932
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3093..3681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3797..4080
                     /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 ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1981 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2041 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2101 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2161 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2221 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2281 tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2341 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2401 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2461 taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt
     2521 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2581 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2641 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2701 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2761 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2821 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2881 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga
     2941 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta
     3001 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3421 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3481 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3541 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3601 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3661 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3721 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3781 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     3841 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     3901 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     3961 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     4021 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     4081 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
