LOCUS       p3xFLAG-CMV-9.        6373 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a neomycin resistance marker for expression and 
            secretion of N-terminally 3xFLAG-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    p3xFLAG-CMV-9.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6373)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6373
                     /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..1052
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     misc_feature    1050..1116
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     polyA_signal    1114..1736
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        1778..2076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2130..2921
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3576..3710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3716..3734)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3748..3764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3772..3788)
                     /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(3796..3826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3841..3862)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4150..4738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4912..5769)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5770..5874)
                     /label=AmpR promoter
     rep_origin      5901..6373
                     /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 acaaagacca tgacggtgat tataaagatc
     1021 atgacatcga ttacaaggat gacgatgaca agcttgcggc cgcgaattca tcgatagatc
     1081 tgatatcggt accagtcgac tctagaggat cccgggtggc atccctgtga cccctcccca
     1141 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     1201 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     1261 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     1321 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     1381 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     1441 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     1501 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     1561 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     1621 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     1681 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     1741 acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1921 atagtcccgc ccctaactcc gcccatcccg cctccgctaa ctccgcccag ttccgcccat
     1981 tctccgcccc atggctgact aatttttttt atttatgcag gccgaggccg cctcggcctc
     2041 tgagctattc cagaagtagt gaggaggctt ttttggagga attgatcagc ttgggatctg
     2101 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2161 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2221 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2281 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     2341 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2401 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2461 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2521 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2581 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2641 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2701 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2761 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2821 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2881 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     2941 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3001 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3061 gcgcggggat ctcatgctgg agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg
     3121 gttcagctgc tgcctgaggc tggacgacct cgcggagttc taccggcagt gcaaatccgt
     3181 cggcatccag gaaaccagca gcggctatcc gcgcatccat gcccccgaac tgcaggagtg
     3241 gggaggcacg atggccgctt tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa
     3301 cgaattgctt gcaggcatct catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg
     3361 tggatggagc gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc
     3421 tggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccg atcataatca
     3481 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3541 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3601 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3661 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aattccctat
     3721 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     3781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4561 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5881 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6181 gtccacgttc tttaagagtc cactattaaa gaacgtggac tcttgttcca aactggaaca
     6241 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6361 acgcttacaa ttt
//
