LOCUS       Exported                8069 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEF6-Etag-mcPLA2, complete sequence.
ACCESSION   LT727058
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  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  JOURNAL   Unpublished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 8069)
  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 8069)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8069
                     /organism="Mammalian expression vector pEF6-Etag-mcPLA2"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4930."
                     /db_xref="taxon:1944996"
     promoter        351..1529
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     regulatory      386..1529
                     /regulatory_class="promoter"
                     /note="hEF1 promoter"
     misc_feature    582..1520
                     /label=intron hEF1 promoter
                     /note="intron hEF1 promoter"
     intron          582..1520
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1546..1564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1546..1562
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    1639..1677
                     /label=E-tag
                     /note="E-tag"
     CDS             1687..3933
                     /codon_start=1
                     /note="unnamed protein product; mcPLA2"
                     /protein_id="SJL87312.1"
                     /translation="MSFIDPYQHIIVEHQYSHKFTVVVLRATKVTKGTFGDMLDTPDPY
                     VELFISTTPDSRKRTRHFNNDINPVWNETFEFILDPNQENVLEITLMDANYVMDETLGT
                     ATFPVSSMKVGEKKEVPFIFNQVTEMILEMSLEVCSCPDLRFSMALCDQEKTFRQQRKE
                     NIKENMKKLLGPKKSEGLYSTRDVPVVAILGSGGGFRAMVGFSGVMKALYESGILDCAT
                     YIAGLSGSTWYMSTLYSHPDFPEKGPEEINEELMKNVSHNPLLLLTPQKVKRYVESLWK
                     KKSSGQPVTFTDIFGMLIGETLIQNRMSMTLSSLKEKVNAARCPLPLFTCLHVKPDVSE
                     LMFADWVEFSPYEIGMAKYGTFMAPDLFGSKFFMGTVVKKYEENPLHFLMGVWGSAFSI
                     LFNRVLGVSGSQNKGSTMEEELENITAKHIVSNDSSDSDDEAQGPKGTENEEAEKEYQS
                     DNQASWVHRMLMALVSDSALFNTREGRAGKVHNFMLGLNLNTSYPLSPLRDFSSQDSFD
                     DELDAAVADPDEFERIYEPLDVKSKKIHVVDSGLTFNLPYPLILRPQRGVDLIISFDFS
                     ARPSDTSPPFKELLLAEKWAKMNKLPFPKIDPYVFDREGLKECYVFKPKNPDVEKDCPT
                     IIHFVLANINFRKYKAPGVLRETKEEKEIADFDIFDDPESPFSTFNFQYPNQAFKRLHD
                     LMYFNTLNNIDVIKDAIVESIEYRRQNPSRCSVSLSNVEARKFFNKEFLSKPTV"
     misc_feature    3951..3980
                     /label=Myc epitope
                     /note="Myc epitope"
     CDS             3951..3980
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3996..4013
                     /label=His-tag
                     /note="His-tag"
     CDS             3996..4013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      4042..4266
                     /regulatory_class="terminator"
                     /note="bovine growth hormone polyA"
     polyA_signal    4042..4266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4312..4740
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      4745..5026
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        4754..5083
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4934..5069
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4983..5058
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      5131..5196
                     /regulatory_class="promoter"
                     /note="synthetic EM7 promoter"
     promoter        5131..5178
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5197..5595
                     /codon_start=1
                     /note="unnamed protein product; blasticidin"
                     /protein_id="SJL87313.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5753..5874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(5753..5827)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      5753..5827
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(5923..5939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5947..5963
                     /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(5966..6049)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(5971..6001)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6016..6037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6141..6532)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(6314..6902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7073..7933)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87314.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7934..8038)
                     /gene="bla"
                     /label=AmpR promoter