LOCUS       pFLAG-CMV-3.        6331 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a neomycin resistance marker for expression and 
            secretion of N-terminally FLAG-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pFLAG-CMV-3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6331)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6331)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     This sequence provided by Sigma-Aldrich may contain errors in the 
            SV40 promoter and f1 ori regions. See pFLAG-CMV(TM)-4 for 
            comparison.
FEATURES             Location/Qualifiers
     source          1..6331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             942..986
                     /label=preprotrypsin leader
                     /note="leader sequence from mouse preprotrypsin"
     CDS             987..1010
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    1008..1074
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     polyA_signal    1072..1694
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        1736..2034
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2088..2879
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3534..3668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3674..3692)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3706..3722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3730..3746)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3754..3784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3799..3820)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4108..4696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4870..5727)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5728..5832)
                     /label=AmpR promoter
     rep_origin      5859..6331
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcag atcgatctac catgtctgca cttctgatcc
      961 tagctcttgt tggagctgca gttgctgact acaaagacga tgacgacaag cttgcggccg
     1021 cgaattcatc gatagatctg atatcggtac cagtcgactc tagaggatcc cgggtggcat
     1081 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     1141 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     1201 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     1261 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     1321 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     1381 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     1441 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     1501 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     1561 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     1621 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     1681 atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt
     1741 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1801 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1861 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc tccgctaact
     1921 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcaggc
     1981 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggaat
     2041 tgatcagctt gggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2101 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     2161 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     2221 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     2281 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     2341 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     2401 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     2461 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     2521 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     2581 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     2641 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     2701 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     2761 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     2821 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     2881 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     2941 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3001 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccgggct
     3061 cgatcccctc gcgagttggt tcagctgctg cctgaggctg gacgacctcg cggagttcta
     3121 ccggcagtgc aaatccgtcg gcatccagga aaccagcagc ggctatccgc gcatccatgc
     3181 ccccgaactg caggagtggg gaggcacgat ggccgctttg gtcgacccgg acgggacgct
     3241 cctgcgcctg atacagaacg aattgcttgc aggcatctca tgagtgtgtc ttcccgtttt
     3301 ccgcctgagg tcactgcgtg gatggagcgc tggcgcctgc tgcgcgacgg cgagctgctc
     3361 accacccact cgccaagctg gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3481 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3541 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3601 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3661 tctggatcaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc
     3721 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3781 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3841 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3901 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3961 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4681 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4801 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4861 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4921 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4981 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5041 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5101 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5161 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5221 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5281 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5341 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5401 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5461 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5521 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5581 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5641 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5701 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5761 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5821 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     5881 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5941 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6001 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6061 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6121 tcgccctttg acgttggagt ccacgttctt taagagtcca ctattaaaga acgtggactc
     6181 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6241 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6301 aattttaaca aaatattaac gcttacaatt t
//
