LOCUS       40924_19081        4594 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin/G418 resistance marker, for protein
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4594)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4594)
  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..4594
                     /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
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     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        1958..2315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2330..2377
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2409..3200
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3267..3315
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3554..4142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4258..4541
                     /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 taagtaagga
     1081 atccacatgg cacaggttat caacacgttt gacggggttg cggattatct tcagacatat
     1141 cataagctac ctgataatta cattacaaaa tcagaagcac aagccctcgg ctgggtggca
     1201 tcaaaaggga accttgcaga cgtcgctccg gggaaaagca tcggcggaga catcttctca
     1261 aacagggaag gcaaactccc gggcaaaagc ggacgaacat ggcgtgaagc ggatattaac
     1321 tatacatcag gcttcagaaa ttcagaccgg attctttact caagcgactg gctgatttac
     1381 aaaacaacgg accattatca gacctttaca aaaatcagat aatgtttaaa cgaattcggg
     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 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     1861 cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg
     1921 aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt
     1981 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2041 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2101 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2161 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2221 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2281 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg ttgacaatta
     2341 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     2401 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2461 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2521 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2581 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2641 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2701 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     2761 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2821 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2881 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     2941 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3001 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3061 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3121 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3181 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3241 gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     3301 ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg
     3361 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     3421 tctacccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3481 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3541 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3601 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3661 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3721 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3781 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3841 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3901 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3961 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4021 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4081 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     4141 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4201 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     4261 gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg
     4321 cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc
     4381 atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt
     4441 ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga
     4501 ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg
     4561 gccacgatgc gtccggcgta gaggatcgag atct
//
