LOCUS       40924_20326        4140 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for mammalian 
            expression from a moderately 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 4140)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4140)
  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..4140
                     /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             143..1033
                     /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             1046..1066
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="EDLYFQS"
     CDS             1104..1436
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    1449..1504
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     polyA_signal    complement(1609..1730)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1977..2081
                     /label=AmpR promoter
     CDS             2082..2939
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3100..3688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3804..4087
                     /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 ggagaattca tcatggcaga aatcggtact ggctttccat tcgaccccca
      181 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac
      241 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc
      301 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga
      361 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga
      421 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt
      481 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg
      541 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg
      601 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct
      661 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt
      721 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg
      781 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc
      841 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc
      901 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct
      961 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtcgacgct
     1021 cgagatttcc ggcgagccaa ccactgagga tctgtacttt cagagcgata acgcgatcgc
     1081 catggaataa gtaaggaatc cacatggcac aggttatcaa cacgtttgac ggggttgcgg
     1141 attatcttca gacatatcat aagctacctg ataattacat tacaaaatca gaagcacaag
     1201 ccctcggctg ggtggcatca aaagggaacc ttgcagacgt cgctccgggg aaaagcatcg
     1261 gcggagacat cttctcaaac agggaaggca aactcccggg caaaagcgga cgaacatggc
     1321 gtgaagcgga tattaactat acatcaggct tcagaaattc agaccggatt ctttactcaa
     1381 gcgactggct gatttacaaa acaacggacc attatcagac ctttacaaaa atcagataat
     1441 gtttaaacga attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca
     1501 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1561 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     1621 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1681 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1741 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     1801 aaaacctcta caaatgtggt aaaatcgaat tctaatggat cctctttgcg cttgcgtttt
     1861 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     1921 tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1981 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2041 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2101 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2161 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2221 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2281 tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2341 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2401 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2461 tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa
     2521 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2581 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2641 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2701 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2761 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2821 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2881 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2941 aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat
     3001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3541 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3661 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     3721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3781 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
     3841 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
     3901 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
     3961 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
     4021 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
     4081 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
