LOCUS       Exported                8321 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-prepro-TSE, complete sequence.
ACCESSION   LT727319
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8321)
  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  JOURNAL   Unpublished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 8321)
  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 8321)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8321
                     /organism="Mammalian expression vector pCAGGS-prepro-TSE"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4172."
                     /db_xref="taxon:1944894"
     CDS             428..694
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=-alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     misc_difference 428..670
                     /label=ppMF
                     /note="ppMF"
     CDS             698..2629
                     /codon_start=1
                     /note="unnamed protein product; mature TS"
                     /protein_id="SJL88095.1"
                     /translation="MLAPGSSRVELFKRKNSTVPFEDKAGKVTERVVHSFRLPALVNVD
                     GVMVAIADARYDTSNDNSLIDTVAKYSVDDGETWETQIAIKNSRVSSVSRVVDPTVIVK
                     GNKLYVLVGSYYSSRSYWSSHGDARDWDILLAVGEVTKSIAGGKITASIKWGSPVSLKK
                     FFPAEMEGMHTNQFLGGAGVAIVASNGNLVYPVQVTNKRKQVFSKIFYSEDDGKTWKFG
                     KGRSDFGCSEPVALEWEGKLIINTRVDWKRRLVYESSDMGNTWVEAVGTLSRVWGPSPK
                     SDHPGSQSSFTAVTIEGMRVMLFTHPLNFKGRWLRDRLNLWLTDNQRIYNVGQVSIGDE
                     NSAYSSVLYKDDKLYCLHEINTDEVYSLVFARLVGELRIIKSVLRSWKNWDSHLSSICT
                     PADPAASSSESGCGPAVTTVGLVGFLSGNASQNVWEDAYRCVNASTANAERVRNGLKFA
                     GVGGGALWPVSQQGQNQRYRFANHAFTLVASVTIHEAPRAASPLLGASLDSSGGKKLLG
                     LSYDEKHQWQPIYGSTPVTPTGSWETGKRYHVVLTVANKIGSVYIDGELLEGSGQTVVP
                     DGRTPDISHFYVGGYGRSDMPTISHVTVNNVLLYNRQLNTEEIRTLFLSQDLIGTEAHM
                     DSSSDTKEL"
     misc_feature    2630..2668
                     /label=E-tag
                     /note="E-tag"
     regulatory      3611..3702
                     /regulatory_class="terminator"
                     /note="3 UTR of rabbit beta-globin, incl. polyA"
     polyA_site      3678..3733
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    3703..4059
                     /label=3 flanking region of rabbit beta-globin
                     /note="3 flanking region of rabbit beta-globin"
     primer_bind     complement(4094..4110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4118..4134
                     /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(4137..4219)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(4142..4172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4186..4207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4266..4461
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4312..4447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4361..4436
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    4467..4601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(4467..4541)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      4467..4541
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      complement(4656..5057)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(4839..5427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5598..6458)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88096.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6459..6563)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      6591..6971
                     /regulatory_class="promoter"
                     /note="hCMV enhancer"
     enhancer        6594..6973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6975..7252
                     /label=chicken beta-actin promoter
     regulatory      6977..8318
                     /regulatory_class="promoter"
                     /note="AG promoter"
     intron          7253..8270
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    7346..8270
                     /label=intron AG promoter
                     /note="intron AG promoter"