LOCUS       Exported                7159 bp ds-DNA    circular SYN 23-2-2013
DEFINITION  Vector for expression of N-terminally FLAG-tagged proteins in 
            bacterial, insect, and mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pDream2.1 MCS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7159)
  AUTHORS   GenScript
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7159
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_recomb     181..1008
                     /note="baculovirus recombination region (lef2/ORF603)"
                     /note="contains ORF603 and part of lef2"
     CDS             181..305
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /note="lef2"
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(343..948)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /note="ORF603"
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1086..1389
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1390..1593
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        1615..1633
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1634..1658
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     promoter        1661..1770
                     /note="p10 promoter"
                     /note="baculovirus promoter for expression in insect cells"
     protein_bind    1772..1796
                     /gene="mutant version of attB"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1810..1812
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             1813..1836
                     /codon_start=1
                     /product="FLAG epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     misc_feature    1837..1882
                     /note="MCS"
                     /note="multiple cloning site"
     protein_bind    1905..1929
                     /gene="mutant version of attB"
                     /bound_moiety="BP clonase(TM)"
                     /note="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1931..2407
                     /note="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     terminator      2408..2455
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2548..2976
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2990..3319
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3170..3305
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3386..4180
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4354..4475
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     polyA_site      4519..4574
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     misc_recomb     4722..5427
                     /note="baculovirus recombination region (ORF1629)"
                     /note="contains part of ORF1629"
     CDS             complement(4839..5427)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /note="ORF1629"
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      complement(5637..6224)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6237..7097)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"