LOCUS       Exported                5471 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-EhRac1-DN, complete sequence.
ACCESSION   LT727151
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5471)
  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  JOURNAL   Unpublished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 5471)
  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 5471)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5471
                     /organism="Mammalian expression vector pCAGGS-EhRac1-DN"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4659."
                     /db_xref="taxon:1944780"
     CDS             4..639
                     /codon_start=1
                     /note="unnamed protein product; hRac1-DN"
                     /protein_id="SJL87607.1"
                     /translation="NSTMGAPVPYPDPLEPRAAAQAIKCVVVGDGAVGKNCLLISYTTN
                     AFPGEYIPTVFDNYSANVMVDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLV
                     SPASFENVRAKWYPEVRHHCPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLA
                     MAKEIGAVKYLECSALTQRGLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     misc_feature    16..54
                     /label=E-tag
                     /note="E-tag"
     CDS             64..639
                     /codon_start=1
                     /product="human Rac family small GTPase 1"
                     /label=Rac1
                     /translation="QAIKCVVVGDGAVGKNCLLISYTTNAFPGEYIPTVFDNYSANVMV
                     DGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHC
                     PNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQRG
                     LKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     CDS             64..639
                     /codon_start=1
                     /product="dominant negative mutant of human Rac family 
                     small GTPase 1"
                     /label=Rac1 (T17N)
                     /translation="QAIKCVVVGDGAVGKNCLLISYTTNAFPGEYIPTVFDNYSANVMV
                     DGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHC
                     PNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQRG
                     LKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     regulatory      761..852
                     /regulatory_class="terminator"
                     /note="beta-globin polyA"
     polyA_site      828..883
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    853..1209
                     /label=3 FR of beta-globin
                     /note="3 FR of beta-globin"
     primer_bind     complement(1244..1260)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1268..1284
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      complement(1287..1369)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(1292..1322)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1336..1357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1416..1611
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1462..1597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      1511..1586
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    1617..1751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(1617..1691)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      1617..1691
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      complement(1806..2207)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(1989..2577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2748..3608)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87608.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3609..3713)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3744..4123
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3744..4121
                     /regulatory_class="promoter"
                     /note="hCMV-IE enhancer"
     promoter        4125..4402
                     /label=chicken beta-actin promoter
     regulatory      4127..5468
                     /regulatory_class="promoter"
                     /note="AG promoter"
     intron          4403..5420
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    4496..5420
                     /label=intron
                     /note="intron"