LOCUS       40924_8231        5468 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-EhRhoA-DN, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5468)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5468)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 5468)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5468)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    16..54
                     /label=E-tag
                     /note="E-tag"
     CDS             64..639
                     /label=RhoA (T19N)
                     /note="dominant negative mutant of human Rho family small
                     GTPase RhoA"
     polyA_signal    825..880
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1241..1257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1265..1281)
                     /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(1289..1319)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1333..1354)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1412..1608
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    1614..1748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1986..2574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2748..3605)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3606..3710)
                     /label=AmpR promoter
     enhancer        3741..4120
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3741..4118
                     /label=hCMV-IE enhancer
                     /note="hCMV-IE enhancer"
                     /regulatory_class="promoter"
     promoter        4122..4399
                     /label=chicken beta-actin promoter
     intron          4400..5417
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
ORIGIN
        1 ttgaattcca ccatgggtgc gccggtgccg tatccggacc cgctggaacc gcgtgcggcc
       61 gctgctgcca tccggaagaa actggtgatt gttggtgatg gagcctgtgg aaagaactgc
      121 ttgctcatag tcttcagcaa ggaccagttc ccagaggtgt atgtgcccac agtgtttgag
      181 aactatgtgg cagatatcga ggtggatgga aagcaggtag agttggcttt gtgggacaca
      241 gctgggcagg aagattatga tcgcctgagg cccctctcct acccagatac cgatgttata
      301 ctgatgtgtt tttccatcga cagccctgat agtttagaaa acatcccaga aaagtggacc
      361 ccagaagtca agcatttctg tcccaacgtg cccatcatcc tggttgggaa taagaaggat
      421 cttcggaatg atgagcacac aaggcgggag ctagccaaga tgaagcagga gccggtgaaa
      481 cctgaagaag gcagagatat ggcaaacagg attggcgctt ttgggtacat ggagtgttca
      541 gcaaagacca aagatggagt gagagaggtt tttgaaatgg ctacgagagc tgctctgcaa
      601 gctagacgtg ggaagaaaaa atctggttgc cttgtcttgt gacccgggac attaaacgcg
      661 tgggtaccgg atccggcaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
      721 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
      781 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
      841 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
      901 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
      961 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     1021 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     1081 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     1141 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     1201 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1261 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1321 cctgggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1381 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     1441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     1621 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1681 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1741 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcaggga taacgcagga
     1921 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1981 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2041 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2101 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2161 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2221 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2281 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2341 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2401 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2461 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2521 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2581 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2641 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2701 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2761 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2821 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2881 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2941 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3001 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3061 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3121 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3181 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3241 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3301 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3361 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3421 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3481 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3541 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3601 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3661 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3721 cgaaaagtgc cacctgggtc gacattgatt attgactagt tattaatagt aatcaattac
     3781 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3841 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3901 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3961 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     4021 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4081 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     4141 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     4201 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggccccccc caggcggggc
     4261 ggggcggggc gaggggcggg gcggggcgag gcggaaaggt gcggcggcag ccaatcagag
     4321 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     4381 gcgaagcgcg cggcgggcgg gagtcgttgc gcgctgcctt ccccccgtgc cccgctccgc
     4441 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     4501 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     4561 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     4621 cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct
     4681 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg
     4741 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa
     4801 aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg
     4861 gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc
     4921 ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt
     4981 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aaggggcgcg
     5041 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     5101 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     5161 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     5221 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     5281 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     5341 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     5401 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     5461 gcaaagaa
//
