LOCUS       Exported                8707 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pDM359, complete sequence.
ACCESSION   EU908846
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8707)
  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 8707)
  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 8707)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8707
                     /organism="Expression vector pDM359"
                     /mol_type="other DNA"
                     /label=Tet-on inducible
                     /note="Tet-on inducible"
                     /db_xref="taxon:554139"
     regulatory      15..446
                     /regulatory_class="promoter"
                     /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..2072)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=hygromycin B phosphotransferase
                     /protein_id="ACG80836.1"
                     /translation="MDQFNISQQDLMKKPELTATSVEKFLIEKFDSVSDLMQLSEGEES
                     RAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISR
                     RAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFIC
                     AIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRI
                     TAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIG
                     LDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSG
                     NRRPSTRPDREMGEAN"
     CDS             complement(1002..2039)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     regulatory      complement(2084..2305)
                     /regulatory_class="promoter"
                     /note="act6 promoter"
     promoter        2360..2464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2465..3325
                     /codon_start=1
                     /product="penicillin beta-lactamase"
                     /label=penicillin beta-lactamase
                     /protein_id="ACG80837.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3473..4140)
                     /direction=LEFT
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      4297..4580
                     /regulatory_class="promoter"
                     /note="act15 promoter"
     CDS             4593..5471
                     /codon_start=1
                     /product="rtTA-M2s*"
                     /label=rtTA-M2s*
                     /note="transactivator"
                     /protein_id="ACG80838.1"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGSHTRRLSTAPPTDVSL
                     GDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMF
                     TDALGIDEYGG"
     regulatory      5510..5771
                     /regulatory_class="terminator"
                     /note="act8 terminator"
     rep_origin      complement(5896..6438)
                     /direction=LEFT
                     /label=Ddp1 ori
                     /note="Ddp1 ori"
     CDS             6348..6941
                     /codon_start=1
                     /product="G5"
                     /label=G5
                     /protein_id="ACG80839.1"
                     /translation="MFNHIKDSGYYATNEEIEIFLESCTLCKEITAQTKRNSYKKRNII
                     NKLPEEEEEEEEEEEEEEEEEEQEEEVEKPTISEEEEEETPAVSEEEKEEEEQEEDKEK
                     DKEKKIEEDTETGKKKDEVMNERQDGIEISEKTNDKPTEKTKVKKNVSRVKDINLHKKG
                     NPITPKKIKNFSQQQLRLASGVSNEKRAIITLKK"
     CDS             complement(6949..7713)
                     /codon_start=1
                     /product="D5"
                     /label=D5
                     /protein_id="ACG80840.1"
                     /translation="MYSTTLLFFLFFFIVSKSSVLQNDQIEVKYFQLLIVMYQKENCVE
                     KSMTGAVIYDECNIHGRVETNSTHALFYDDIETNNSRCNNFRNLTNLIKLNECINDEFG
                     ESILYKEYNETDDGYLFRVEDSFVEITSLSMDCTKNSKTIIEKFNICSKFENVYHITNI
                     TQEKSNRFTCTDPLCHYCKNENIQNNLDFKTTKCTPKYGASDSEFLSTIYNPKLDGSNN
                     GMEKSVTQEKNISNNLKINIYLIFFLIIFLIK"
     CDS             complement(8216..8707)
                     /codon_start=1
                     /product="D1"
                     /label=D1
                     /protein_id="ACG80841.1"
                     /translation="MDFEKVNSNYYKIQSESTQINSNEIADLKKFIKEEVNKTSSKIDF
                     FLVSSTDALSNPENYSLLEVKCINCHSLCQGKNLYISCTRDGCQNNICYNCLGININIY
                     NVVINSKLCPPCFNDSVINKKCAMCSKNGTKCNLNQECKLHLCAQCSKKCLYILRVKTN
                     "