LOCUS       Exported                6734 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCR3VSV-CED-4, complete sequence.
ACCESSION   LT727395
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6734)
  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  JOURNAL   Unpublished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 6734)
  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 6734)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6734
                     /organism="Mammalian expression vector pCR3VSV-CED-4"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3811."
                     /db_xref="taxon:1944964"
     regulatory      10..593
                     /regulatory_class="promoter"
                     /note="hCMV promoter"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      638..654
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_difference 691..723
                     /label=VSV epitope
                     /note="VSV epitope"
     CDS             691..723
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             736..2385
                     /codon_start=1
                     /note="unnamed protein product; CED-4"
                     /protein_id="SJL88336.1"
                     /translation="MLCEIECRALSTAHTRLIHDFEPRDALTYLEGKNIFTEDHSELIS
                     KMSTRLERIANFLRIYRRQASELGPLIDFFNYNNQSHLADFLEDYIDFAINEPDLLRPV
                     VIAPQFSRQMLDRKLLLGNVPKQMTCYIREYHVDRVIKKLDEMCDLDSFFLFLHGRAGS
                     GKSVIASQALSKSDQLIGINYDSIVWLKDSGTAPKSTFDLFTDILLMLKSEDDLLNFPS
                     VEHVTSVVLKRMICNALIDRPNTLFVFDDVVQEETIRWAQELRLRCLVTTRDVEISNAA
                     SQTCEFIEVTSLEIDECYDFLEAYGMPMPVGEKEEDVLNKTIELSSGNPATLMMFFKSC
                     EPKTFEKMAQLNNKLESRGLVGVECITPYSYKSLAMALQRCVEVLSDEDRSALAFAVVM
                     PPGVDIPVKLWSCVIPVDICSNEEEQLDDEVADRLKRLSKRGALLSGKRMPVLTFKIDH
                     IIHMFLKHVVDAQTIANGISILEQRLLEIGNNNVSVPERHIPSHFQKFRRSSASEMYPK
                     TTEETVIRPEDFPKFMQLHQKFYDSLKNFACC"
     promoter        complement(2449..2467)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(2451..2467)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     regulatory      2493..2717
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    2493..2717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2741..3067)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(2849..3437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    3766..3813
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      complement(3785..3854)
                     /regulatory_class="terminator"
                     /note="TK polyA"
     CDS             complement(4045..4833)
                     /codon_start=1
                     /note="unnamed protein product; neomycin resistance"
                     /protein_id="SJL88337.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4868..5225)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4882..5017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(4893..4968)
                     /direction=LEFT
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      complement(4925..5206)
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     CDS             complement(5285..6145)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88338.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6146..6250)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6277..6732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"