LOCUS       p3xFLAG-CMV-8.        4763 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for expression and secretion of N-terminally 3xFLAG-tagged 
            proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    p3xFLAG-CMV-8.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4763)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4763)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4763
                     /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             932..976
                     /label=preprotrypsin leader
                     /note="leader sequence from mouse preprotrypsin"
     CDS             977..1042
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     misc_feature    1040..1106
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     polyA_signal    1104..1726
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        1755..2084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        complement(2123..2141)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2155..2171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2179..2195)
                     /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(2203..2233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2248..2269)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2557..3145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3319..4176)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4177..4281)
                     /label=AmpR promoter
     rep_origin      4308..4763
                     /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 attaattcac catgtctgca cttctgatcc tagctcttgt
      961 tggagctgca gttgctgact acaaagacca tgacggtgat tataaagatc atgacatcga
     1021 ttacaaggat gacgatgaca agcttgcggc cgcgaattca tcgatagatc tgatatcggt
     1081 accagtcgac tctagaggat cccgggtggc atccctgtga cccctcccca gtgcctctcc
     1141 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     1201 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     1261 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     1321 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     1381 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     1441 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     1501 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     1561 ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga
     1621 cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca
     1681 ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag
     1741 cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ctcgaggaac
     2101 tgaaaaacca gaaagttaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata
     2161 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2221 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2281 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3301 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3361 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3421 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3481 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3541 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3601 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3661 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3721 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3781 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3841 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3901 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3961 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4021 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4081 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4141 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     4321 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4381 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4441 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4501 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4561 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4621 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4681 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4741 caaaatatta acgcttacaa ttt
//
