LOCUS       40924_19066        4123 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for protein 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4123)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4123)
  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..4123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1087..1419
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLQGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    1432..1487
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     polyA_signal    complement(1592..1713)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1960..2064
                     /label=AmpR promoter
     CDS             2065..2922
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3083..3671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3787..4070
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatggaa taagtaaagg
     1081 aatcacatgg cacaggttat caacacgttt gacggggttg cggattatct tcagacatat
     1141 cataagctac ctgataatta cattacaaaa tcagaagcac aagccctcgg ctgggtggca
     1201 tcaaaaggga accttgcaga cgtcgctccg gggaaaagca tcggcggaga catcttctca
     1261 aacagggaag gcaaactcca gggcaaaagc ggacgaacat ggcgtgaagc ggatattaac
     1321 tatacatcag gcttcagaaa ttcagaccgg attctttact caagcgactg gctgatttac
     1381 aaaacaacgg accattatca gacctttaca aaaatcagat aatgtttaaa cgaattcgag
     1441 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta
     1501 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1561 cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     1621 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1681 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1741 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     1801 ggtaaaatcg aattctaatg gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc
     1861 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtaatg
     1921 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1981 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2041 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2101 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2161 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2221 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt
     2281 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2341 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2401 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2461 gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2521 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2581 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2641 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2701 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2761 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2821 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2881 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga aatgaccgac
     2941 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3481 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3661 aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3781 tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat
     3841 gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca
     3901 taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg
     3961 ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca
     4021 attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc
     4081 agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
