LOCUS       sequence-319252-        7701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector optimized for use in screening assays, as well as for 
            efficient production of baculovirus and robust expression of the 
            target protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7701)
  AUTHORS   Goehring A, Lee CH, Wang KH, Michel JC, Claxton DP, Baconguis I, 
            Althoff T, Fischer S, Garcia KC, Gouaux E
  TITLE     Screening and large-scale expression of membrane proteins in 
            mammalian cells for structural studies.
  JOURNAL   Nat Protoc. 2014 Nov;9(11):2574-85. doi: 10.1038/nprot.2014.173. 
            Epub 2014 Oct 9.
  PUBMED    25299155
REFERENCE   2  (bases 1 to 7701)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nprot.2014.173"; journalName: "Nat Protoc"; date: 
            "2014-11"; volume: "9"; issue: "11"; pages: "2574-85"
COMMENT     Sequence Label: pEG BacMam N term myc eGFP 3C
FEATURES             Location/Qualifiers
     source          1..7701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        180..559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        560..763
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     714..734
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     760..784
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          899..1031
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1014..1033
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1100..1129
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1139..1141,1142..1852)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1181..1202)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1442..1461)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1789..1810
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1871..1894
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    1972..2560
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2025..2045)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2443..2454)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2693..2827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2730..2749)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2784..2803
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2856..3021
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3205..3659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3292..3311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3501..3522
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3685..3789
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3790..3858,3859..4650)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4008..4027)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4821..5409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5310..5329
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5563..5580
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5714..5938
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6005..6538)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6727..6732,6733..6749,6750..6755))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7272..7320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     7345..7364
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(7456..7565)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        7610..7701
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7626..7643
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cctcaatatt ggccattagc
       61 catattattc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
      121 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
      181 gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      241 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      301 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      361 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      421 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      481 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
      541 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
      601 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      661 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
      721 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
      781 gatcactaga agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc
      841 ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
      901 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
      961 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1021 tctctccaca ggtgtccact cccagttcaa ttacagctct taagggatcc cggtccgaag
     1081 cgcgcggaat tacaccatgg aacaaaaact catctcagaa gaggatctgg gttcaggagt
     1141 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1201 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1261 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1321 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1381 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1441 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1501 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1561 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1621 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1681 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1741 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1801 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     1861 cagatctgga ctggaagttc tgttccaggg tcctgaattc aaaggcctac gtcgacgagc
     1921 tcactagtcg cggccgcttt cgaatctaga gcctgcagtc tcgacaagct taatcaacct
     1981 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2041 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2101 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2161 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2221 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2281 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2341 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2401 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2461 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2521 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgaagcttg tcgagaagta
     2581 ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2641 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2701 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2761 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2821 ctggatctga tcactgcttg agcctaggag atccgaacca gataagtgaa atctagttcc
     2881 aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc
     2941 tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc
     3001 caactatttt gtccgcccac agcggggcat ttttcttcct gttatgtttt taatcaaaca
     3061 tcctgccaac tccatgtgac aaaccgtcat cttcggctac tttttctctg tcacagaatg
     3121 aaaatttttc tgtcatctct tcgttattaa tgtttgtaat tgactgaata tcaacgctta
     3181 tttgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3361 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3481 tggagtccac gttcttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3541 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3601 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3661 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3721 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3781 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3841 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3901 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3961 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4021 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4081 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4141 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4201 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4261 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4321 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4381 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4441 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4501 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4561 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4621 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4681 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4741 aatctcatga ccaaaatccc ttaacatgag ttttcgttcc actgagcgtc agaccccgta
     4801 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4861 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4921 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4981 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5041 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5101 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5161 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5221 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5281 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5341 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5401 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5461 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5521 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5581 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5641 cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatgcc
     5701 ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact gaacaaaata gatctaaact
     5761 atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt ccagttatgc
     5821 tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt ttctgaagtg
     5881 caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga ctatattcgc
     5941 ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga
     6001 attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca
     6061 actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat
     6121 aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct
     6181 gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct
     6241 caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag
     6301 caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat
     6361 gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca
     6421 tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac
     6481 ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg
     6541 ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc
     6601 cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg
     6661 ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgatcgcat acgctacttg
     6721 cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcctt catccgtttc
     6781 cacggtgtgc gtcacccggc aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg
     6841 ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc ggccttgctg
     6901 ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc ttcaggagat cggtagacct
     6961 cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg aagtggttcg catcctcggt
     7021 tttctggaag gcgagcatcg tttgttcgcc caggactcta gctatagttc tagtggttgg
     7081 ctacagcttt gtttaaacaa agctggctat ggcagggctt gccgccccga cgttggctgc
     7141 gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag aaacgcgggc
     7201 gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     7261 cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt tttattgccg
     7321 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctcccccatc
     7381 tcccggtacc gcatgctatg catcagctgc tagcaccatg gctcgagatc ccgggtgatc
     7441 aagtcttcgt cgagtgattg taaataaaat gtaatttaca gtatagtatt ttaattaata
     7501 tacaaatgat ttgataataa ttcttattta actataatat attgtgttgg gttgaattaa
     7561 aggtccgtat actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
     7621 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     7681 gtaataaaaa aacctataaa t
//
