LOCUS       pFLAG-CMV-4.        6271 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a neomycin resistance marker for expression of 
            N-terminally FLAG-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pFLAG-CMV-4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6271)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6271)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6271
                     /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..944
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             945..968
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    966..1032
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     polyA_signal    1030..1652
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        1694..1991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2045..2836
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3491..3625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3631..3649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3663..3679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3687..3703)
                     /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(3711..3741)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3756..3777)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4065..4653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4827..5684)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5685..5789)
                     /label=AmpR promoter
     rep_origin      5816..6271
                     /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 catggactac aaagacgatg
      961 acgacaagct tgcggccgcg aattcatcga tagatctgat atcggtacca gtcgactcta
     1021 gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     1081 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     1141 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     1201 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     1261 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     1321 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     1381 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     1441 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     1501 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     1561 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     1621 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat
     1681 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1741 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     1801 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     1861 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     1921 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     1981 ggcttttttg gaggaattga tcagcttggg atctgatcaa gagacaggat gaggatcgtt
     2041 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2101 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2161 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2221 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2281 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2341 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     2401 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     2461 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     2521 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     2581 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     2641 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2701 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     2761 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2821 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     2881 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     2941 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3001 ttcgcccacc ccgggctcga tcccctcgcg agttggttca gctgctgcct gaggctggac
     3061 gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca tccaggaaac cagcagcggc
     3121 tatccgcgca tccatgcccc cgaactgcag gagtggggag gcacgatggc cgctttggtc
     3181 gacccggacg ggacgctcct gcgcctgata cagaacgaat tgcttgcagg catctcatga
     3241 gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat ggagcgctgg cgcctgctgc
     3301 gcgacggcga gctgctcacc acccactcgc caagctggaa ccgtaaaaag gccgcgttgc
     3361 tggcgttttt ccataggctc cgccgatcat aatcagccat accacatttg tagaggtttt
     3421 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3481 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3601 caatgtatct tatcatgtct ggatcaattc cctatagtga gtcgtattaa attcgtaatc
     3661 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3961 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4021 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4081 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4141 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4201 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4261 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4321 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4381 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4441 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4501 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4561 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4621 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4681 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4741 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4801 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4861 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4921 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4981 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5041 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5101 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5161 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5221 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5281 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5341 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5401 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5461 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5521 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5581 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5641 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5701 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5761 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     5821 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5881 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5941 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6001 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6061 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6121 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6181 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6241 aattttaaca aaatattaac gcttacaatt t
//
