LOCUS       Exported                7278 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGSEhA20, complete sequence.
ACCESSION   LT727385
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7278)
  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  JOURNAL   Unpublished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 7278)
  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 7278)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7278
                     /organism="Mammalian expression vector pCAGGSEhA20"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3778."
                     /db_xref="taxon:1944763"
     misc_feature    16..54
                     /label=E-tag
                     /note="E-tag"
     CDS             64..2436
                     /codon_start=1
                     /note="unnamed protein product; hA20"
                     /protein_id="SJL88311.1"
                     /translation="MAEQVLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIHHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRNIQATLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHATSQYMWGVQDTDLVLRKALFSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLIKMASTDTPMARSGLQYNSLEEIHIFVLCNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSHHFVPLVTLKDSGPEIRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLMVIEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSEQGRREGHAQNPMEPSVPQLSLMDVKCETPNCPFFMSVN
                     TQPLCHECSERRQKNQNKLPKLNSKPGPEGLPGMALGASRGEAYEPLAWNPEESTGGPH
                     SAPPTAPSPFLFSETTAMKCRSPGCPFTLNVQHNGFCERCHNARQLHASHAPDHTRHLD
                     PGKCQACLQDVTRTFNGICSTCFKRTTAEASSSLSTSLPPSCHQRSKSDPSRLVRSPSP
                     HSCHRAGNDAPAGCLSQAARTPGDRTGTSKCRKAGCVYFGTPENKGFCTLCFIEYRENK
                     HFAAASGKVSPTASRFQNTIPCLGRECGTLGSTMFEGYCQKCFIEAQNQRFHEAKRTEE
                     QLRSSQRRDVPRTTQSTSRPKCARASCKNILACRSEELCMECQHPNQRMGPGAHRGEPA
                     PEDPPKQRCRAPACDHFGNAKCNGYCNECFQFKQMYG"
     misc_feature    2437..2480
                     /label=3 UTR of hA20
                     /note="3 UTR of hA20"
     regulatory      2568..2659
                     /regulatory_class="terminator"
                     /note="3 UTR of rabbit beta-globin, incl. polyA"
     polyA_site      2635..2690
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    2660..3016
                     /label=3 flanking region of rabbit beta-globin
                     /note="3 flanking region of rabbit beta-globin"
     primer_bind     complement(3051..3067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3075..3091
                     /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(3094..3176)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(3099..3129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3143..3164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3223..3418
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3269..3404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3318..3393
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    3424..3558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3424..3498)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3424..3498
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      complement(3613..4014)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3796..4384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4555..5415)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88312.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5416..5520)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5551..5930
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      5551..5928
                     /regulatory_class="promoter"
                     /note="hCMV-IE enhancer"
     promoter        5932..6209
                     /label=chicken beta-actin promoter
     regulatory      5934..7275
                     /regulatory_class="promoter"
                     /note="AG promoter"
     intron          6210..7227
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    6303..7227
                     /label=intron AG promoter
                     /note="intron AG promoter"