LOCUS       40924_19076        5591 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a ampicillin and G418 resistance markers, for 
            protein expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5591)
  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..5591
                     /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"
     CDS             2366..3157
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3224..3272
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        3428..3532
                     /label=AmpR promoter
     CDS             3533..4390
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(4551..5139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5255..5538
                     /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 tgattcttct gacacaacag
     2341 tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc
     2401 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     2461 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     2521 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     2581 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     2641 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     2701 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     2761 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     2821 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     2881 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     2941 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3001 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3061 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3121 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     3181 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     3241 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tcctctttgc
     3301 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     3361 tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca cttttcgggg
     3421 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3481 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3541 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3601 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3661 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3721 ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat cccgtattga
     3781 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3841 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3901 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta tcggaggacc
     3961 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4021 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4081 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4141 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4201 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4261 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4321 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4381 taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     4441 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4501 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4741 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4921 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4981 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5041 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5101 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     5161 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5221 tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac gcacgctgga
     5281 tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga ttaagtgggg
     5341 gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat ttatcaggcg
     5401 cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct gccgtgatcg
     5461 cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc
     5521 tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag
     5581 gatcgagatc t
//
