LOCUS       p3xFLAG-CMV-13.        6362 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a neomycin resistance marker for expression and 
            secretion of C-terminally 3xFLAG-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    p3xFLAG-CMV-13
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6362)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6362
                     /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             931..975
                     /label=preprotrypsin leader
                     /note="leader sequence from mouse preprotrypsin"
     misc_feature    976..1042
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1045..1110
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     polyA_signal    1121..1743
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        1785..2082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2136..2927
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3582..3716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3722..3740)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3754..3770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3778..3794)
                     /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(3802..3832)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3847..3868)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4918..5775)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5776..5880)
                     /label=AmpR promoter
     rep_origin      5907..6362
                     /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 attacggacc atgtctgcac ttctgatcct agctcttgtt
      961 ggagctgcag ttgctaagct tgcggccgcg aattcatcga tagatctgat atcggtacca
     1021 gtcgactcta gaggatcccg ggctgactac aaagaccatg acggtgatta taaagatcat
     1081 gacatcgact acaaggatga cgatgacaag tagtgatccc gggtggcatc cctgtgaccc
     1141 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     1201 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     1261 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     1321 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     1381 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     1441 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     1501 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     1561 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca
     1621 gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt
     1681 gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga
     1741 attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     1801 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     1861 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1921 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     1981 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2041 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggaattg atcagcttgg
     2101 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2161 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2221 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2281 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2341 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2401 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2461 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2521 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2581 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2641 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2701 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     2761 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     2821 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     2881 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     2941 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3001 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3061 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccgggctcg atcccctcgc
     3121 gagttggttc agctgctgcc tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa
     3181 atccgtcggc atccaggaaa ccagcagcgg ctatccgcgc atccatgccc ccgaactgca
     3241 ggagtgggga ggcacgatgg ccgctttggt cgacccggac gggacgctcc tgcgcctgat
     3301 acagaacgaa ttgcttgcag gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc
     3361 actgcgtgga tggagcgctg gcgcctgctg cgcgacggcg agctgctcac cacccactcg
     3421 ccaagctgga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgccgatca
     3481 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3541 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaatt
     3721 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3781 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3841 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3901 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3961 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4981 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5041 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5101 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5161 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5221 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5281 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5341 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5401 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5461 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5521 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5581 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5641 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5701 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5761 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5821 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5881 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5941 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6001 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6061 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6121 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6181 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6241 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6301 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     6361 tt
//
