LOCUS       Exported                8066 bp ds-DNA    circular SYN 22-4-2014
DEFINITION  Baculovirus transfer plasmid encoding beta-glucuronidase, with two 
            polyhedrin and two p10 promoters, for the simultaneous expression of
            up to four proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pBACgus4x-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8066
                     /organism="synthetic DNA construct"
                     /lab_host="Insect Cells"
                     /mol_type="other DNA"
     misc_recomb     329..1156
                     /note="baculovirus recombination region (lef2/ORF603)"
                     /note="contains ORF603 and part of lef2"
     CDS             329..453
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /note="lef2"
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(491..1096)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /note="ORF603"
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     CDS             complement(1167..2978)
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /note="GUS"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFSLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     promoter        complement(2987..3302)
                     /gene="Autographa californica p6.9"
                     /note="p6.9 promoter"
                     /note="baculovirus late basic protein promoter"
     misc_feature    3320..3368
                     /note="poly(A) signal"
                     /note="synthetic polyadenylation signal"
     misc_feature    3371..3397
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(3400..3509)
                     /note="p10 promoter"
                     /note="baculovirus promoter for expression in insect cells"
     misc_feature    3642..3652
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(3660..3751)
                     /gene="polh from Autographa californica"
                     /note="polyhedrin promoter"
                     /note="promoter for the baculovirus polyhedrin gene"
     promoter        3776..3885
                     /note="p10 promoter"
                     /note="baculovirus promoter for expression in insect cells"
     misc_feature    3893..3904
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    3905..3953
                     /note="poly(A) signal"
                     /note="synthetic polyadenylation signal"
     promoter        3991..4082
                     /gene="polh from Autographa californica"
                     /note="polyhedrin promoter"
                     /note="promoter for the baculovirus polyhedrin gene"
     misc_feature    4086..4115
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             4116..4133
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_recomb     4170..5527
                     /note="baculovirus recombination region (ORF1629)"
                     /note="contains part of ORF1629"
     CDS             complement(4186..5527)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /note="ORF1629"
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      complement(5790..6378)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6549..7409)
                     /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"
     promoter        complement(7410..7514)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      7541..7996
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"