LOCUS       Exported                7476 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pcDNA3-hIRAK1-T66A, complete sequence.
ACCESSION   LT726935
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  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  JOURNAL   Unpublished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 7476)
  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 7476)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7476
                     /organism="Mammalian expression vector pcDNA3-hIRAK1-T66A"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5628."
                     /db_xref="taxon:1944933"
     regulatory      235..818
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      863..879
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     CDS             947..3085
                     /codon_start=1
                     /note="unnamed protein product; hIRAK1 mutant"
                     /protein_id="SJL87001.1"
                     /translation="MAGGPGPGEPAAPGAQHFLYEVPPWVMCRFYKVMDALEPADWCQF
                     AALIVRDQTELRLCERSGQRAASVLWPWINRNARVADLVHILTHLQLLRARDIITAWHP
                     PAPLPSPGTTAPRPSSIPAPAEAEAWSPRKLPSSASTFLSPAFPGSQTHSGPELGLVPS
                     PASLWPPPPSPAPSSTKPGPESSVSLLQGARPSPFCWPLCEISRGTHNFSEELKIGEGG
                     FGCVYRAVMRNTVYAVKRLKENADLEWTAVKQSFLTEVEQLSRFRHPNIVDFAGYCAQN
                     GFYCLVYGFLPNGSLEDRLHCQTQACPPLSWPQRLDILLGTARAIQFLHQDSPSLIHGD
                     IKSSNVLLDERLTPKLGDFGLARFSRFAGSSPSQSSMVARTQTVRGTLAYLPEEYIKTG
                     RLAVDTDTFSFGVVVLETLAGQRAVKTHGARTKYLKDLVEEEAEEAGVALRSTQSTLQA
                     GLAADAWAAPIAMQIYKKHLDPRPGPCPPELGLGLGQLACCCLHRRAKRRPPMTQVYER
                     LEKLQAVVAGVPGHLEAASCIPPSPQENSYVSSTGRAHSGAAPWQPLAAPSGASAQAAE
                     QLQRGPNQPVESDESLGGLSAALRSWHLTPSCPLDPAPLREAGCPQGDTAGESSWGSGP
                     GSRPTAVEGLALGSSASSSSEPPQIIINPARQKMVQKLALYEDGALDSLQLLSSSSLPG
                     LGLEQDRQGPEESDEFQS"
     misc_feature    1142..1144
                     /label=T66A
                     /note="T66A"
     promoter        complement(3101..3119)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(3103..3119)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     regulatory      3145..3369
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    3145..3369
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3415..3843
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3848..4057
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3896..4106
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3918..4114
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3965..4100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4014..4089
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             4181..4975
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87002.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5149..5270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(5149..5223)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      5149..5223
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(5319..5335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5343..5359
                     /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(5362..5445)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(5367..5397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5412..5433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5537..5939)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(5721..6309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6480..7340)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87003.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7341..7445)
                     /gene="bla"
                     /label=AmpR promoter