LOCUS       Exported                6640 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-FLAGmA20d5, complete sequence.
ACCESSION   LT727048
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  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  JOURNAL   Unpublished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 6640)
  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 6640)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6640
                     /organism="Mammalian expression vector pCAGGS-FLAGmA20d5"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4844."
                     /db_xref="taxon:1944853"
     misc_feature    13..36
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             13..36
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             46..1914
                     /codon_start=1
                     /note="unnamed protein product; mA20 (d547-699)"
                     /protein_id="SJL87289.1"
                     /translation="MAEQLLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIYHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRSVQASLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHAACQYMWGVQDTDLVLRKALCSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLVKMASADTPAARSGLQYNSLEEIHIFVLSNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSQHFVPLVTLKDSGPELRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLIVMEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSDQARRAAHAQNPLEPSTPQLSLMDIKCETPNCPFFMSVN
                     TQPLCHECSERRQKNQSKLPKLNSKLGPEGLPGVGLGSSNWSPEETAGGPHSAPPTAPS
                     LFLFSETTAMKCRSPGCPFTLNVQHNGFCERCHARQINASHTADPGKCQACLQDVTRTF
                     NGICSTCFKRTTAEPSSSLTSSIPASCHQKCARASCKNILACRSEELCMECQHLSQRVG
                     SVAHRGEPTPEEPPKQRCRAPACDHFGNAKCNGYCNECYQFKQMYG"
     regulatory      1926..2017
                     /regulatory_class="terminator"
                     /note="3 UTR of rabbit beta-globin, incl. polyA"
     polyA_site      1993..2048
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    2018..2374
                     /label=3 flanking region of rabbit beta-globin
                     /note="3 flanking region of rabbit beta-globin"
     primer_bind     complement(2409..2425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2433..2449
                     /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(2452..2534)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(2457..2487)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2501..2522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2581..2776
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2627..2762
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2676..2751
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    2782..2916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(2782..2856)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      2782..2856
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      complement(2971..3372)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3154..3742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3913..4773)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87290.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4774..4878)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4913..5292
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      4913..5290
                     /regulatory_class="promoter"
                     /note="hCMV-IE enhancer"
     promoter        5294..5571
                     /label=chicken beta-actin promoter
     regulatory      5296..6637
                     /regulatory_class="promoter"
                     /note="AG promoter"
     intron          5572..6589
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    5665..6589
                     /label=intron AG promoter
                     /note="intron AG promoter"