LOCUS       Exported                8596 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pDM309, complete sequence.
ACCESSION   EU908848
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8596)
  AUTHORS   Veltman DM, Keizer-Gunnink I, Haastert PJ.
  TITLE     An extrachromosomal, inducible expression system for Dictyostelium 
            discoideum
  JOURNAL   Plasmid 61 (2), 119-125 (2009)
  PUBMED    19046986
REFERENCE   2  (bases 1 to 8596)
  AUTHORS   Veltman DM, Keizer I, van Haastert PJM.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUL-2008) Cell Biochemistry, Rijks Universiteit 
            Groningen, Kerklaan 30, Haren, Groningen 9751 NN, The Netherlands
REFERENCE   3  (bases 1 to 8596)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8596
                     /organism="Expression vector pDM309"
                     /mol_type="other DNA"
                     /label=Tet-off inducible
                     /note="Tet-off inducible"
                     /db_xref="taxon:554137"
     regulatory      15..446
                     /regulatory_class="terminator"
                     /note="TRE-Pmin"
     protein_bind    28..298
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     regulatory      473..734
                     /regulatory_class="terminator"
                     /note="act8 terminator"
     regulatory      complement(741..995)
                     /regulatory_class="terminator"
                     /note="mhcA terminator"
     CDS             complement(1002..1832)
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /protein_id="ACG80848.1"
                     /translation="MDGEDVQAGSFRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSD
                     AAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWL
                     LLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAG
                     LVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCG
                     RLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1002..1796)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(1844..2065)
                     /regulatory_class="promoter"
                     /note="act6 promoter"
     promoter        2120..2224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2225..3085
                     /codon_start=1
                     /product="penicillin beta-lactamase"
                     /label=penicillin beta-lactamase
                     /protein_id="ACG80849.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3233..3900)
                     /direction=LEFT
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      3256..3844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      4057..4340
                     /regulatory_class="promoter"
                     /note="act15 promoter"
     CDS             4353..5360
                     /codon_start=1
                     /product="tTA"
                     /label=tTA
                     /note="transactivator"
                     /protein_id="ACG80850.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     regulatory      5399..5660
                     /regulatory_class="terminator"
                     /note="act8 terminator"
     rep_origin      complement(5785..6327)
                     /direction=LEFT
                     /label=Ddp1 ori
                     /note="Ddp1 ori"
     CDS             6237..6830
                     /codon_start=1
                     /product="G5"
                     /label=G5
                     /protein_id="ACG80851.1"
                     /translation="MFNHIKDSGYYATNEEIEIFLESCTLCKEITAQTKRNSYKKRNII
                     NKLPEEEEEEEEEEEEEEEEEEQEEEVEKPTISEEEEEETPAVSEEEKEEEEQEEDKEK
                     DKEKKIEEDTETGKKKDEVMNERQDGIEISEKTNDKPTEKTKVKKNVSRVKDINLHKKG
                     NPITPKKIKNFSQQQLRLASGVSNEKRAIITLKK"
     CDS             complement(6838..7602)
                     /codon_start=1
                     /product="D5"
                     /label=D5
                     /protein_id="ACG80852.1"
                     /translation="MYSTTLLFFLFFFIVSKSSVLQNDQIEVKYFQLLIVMYQKENCVE
                     KSMTGAVIYDECNIHGRVETNSTHALFYDDIETNNSRCNNFRNLTNLIKLNECINDEFG
                     ESILYKEYNETDDGYLFRVEDSFVEITSLSMDCTKNSKTIIEKFNICSKFENVYHITNI
                     TQEKSNRFTCTDPLCHYCKNENIQNNLDFKTTKCTPKYGASDSEFLSTIYNPKLDGSNN
                     GMEKSVTQEKNISNNLKINIYLIFFLIIFLIK"
     CDS             complement(8105..8596)
                     /codon_start=1
                     /product="D1"
                     /label=D1
                     /protein_id="ACG80853.1"
                     /translation="MDFEKVNSNYYKIQSESTQINSNEIADLKKFIKEEVNKTSSKIDF
                     FLVSSTDALSNPENYSLLEVKCINCHSLCQGKNLYISCTRDGCQNNICYNCLGININIY
                     NVVINSKLCPPCFNDSVINKKCAMCSKNGTKCNLNQECKLHLCAQCSKKCLYILRVKTN
                     "