LOCUS       Exported                5527 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-E-hTRAM, complete sequence.
ACCESSION   LT726979
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5527)
  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  JOURNAL   Unpublished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 5527)
  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 5527)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5527
                     /organism="Mammalian expression vector pCAGGS-E-hTRAM"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5227."
                     /db_xref="taxon:1944784"
     misc_feature    16..54
                     /label=E-tag
                     /note="E-tag"
     CDS             64..771
                     /codon_start=1
                     /note="unnamed protein product; hTRAM"
                     /protein_id="SJL87105.1"
                     /translation="MGIGKSKINSCPLSLSWGKRHSVDTSPGYHESDSKKSEDLSLCNV
                     AEHSNTTEGPTGKQEGAQSVEEMFEEEAEEEVFLKFVILHAEDDTDEALRVQNLLQDDF
                     GIKPGIIFAEMPCGRQHLQNLDDAVNGSAWTILLLTENFLRDTWCNFQFYTSLMNSVNR
                     QHKYNSVIPMRPLNNPLPRERTPFALQTINALEEESRGFPTQVERIFQESVYKTQQTIW
                     KETRNMVQRQFIA"
     regulatory      813..904
                     /regulatory_class="terminator"
                     /note="beta-globin polyA"
     polyA_site      880..935
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    905..1261
                     /label=3 FR of beta-globin
                     /note="3 FR of beta-globin"
     primer_bind     complement(1296..1312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1320..1336
                     /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(1339..1421)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(1344..1374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1388..1409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1468..1663
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1514..1649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      1563..1638
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    1669..1803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(1669..1743)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      1669..1743
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      complement(1858..2259)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(2041..2629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2800..3660)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87106.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3661..3765)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3796..4175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3796..4173
                     /regulatory_class="promoter"
                     /note="hCMV-IE enhancer"
     promoter        4177..4454
                     /label=chicken beta-actin promoter
     regulatory      4179..5524
                     /regulatory_class="promoter"
                     /note="AG promoter"
     misc_feature    4548..5476
                     /label=intron
                     /note="intron"