LOCUS       sequence-319258-        7690 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 7690)
  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 7690)
  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 C term 3C eGFP StrepII
FEATURES             Location/Qualifiers
     source          1..7690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          340..472
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     455..474
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             584..607
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(620..622,623..1333)
                     /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(662..683)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(923..942)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1270..1291
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1349..1372
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1402..1990
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1455..1475)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1873..1884)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2123..2257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2160..2179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2214..2233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2286..2451
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2635..3089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2722..2741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2931..2952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3115..3219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3220..3288,3289..4080)
                     /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(3438..3457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4251..4839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4740..4759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4993..5010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5144..5368
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5435..5968)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6157..6162,6163..6179,6180..6185))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    6702..6750
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6775..6794
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(6886..6995)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        7040..7131
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7056..7073
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     enhancer        7311..7690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcactag aagctttatt
      241 gcggtagttt atcacagtta aattgctaac gcagtcagtg cttctgacac aacagtctcg
      301 aacttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga
      361 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
      421 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac
      481 tcccagttca attacagctc ttaagggatc ccggtccgaa gcgcgcggaa ttcaaaggcc
      541 tacgtcgacg agctcactag tcgcggccgc tttcgaatct agactggaag ttctgttcca
      601 gggtcctgca gccgccgcgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      661 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      721 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      781 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      841 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      901 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      961 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1021 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1081 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1141 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1201 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     1261 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1321 cgagctgtac aagtccggag gttcaggatg gagccacccg cagttcgaaa aatgactaga
     1381 gcctgcagtc tcgacaagct taatcaacct ctggattaca aaatttgtga aagattgact
     1441 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1501 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     1561 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     1621 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     1681 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     1741 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     1801 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     1861 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     1921 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     1981 gcctccccgc ctgaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     2041 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2101 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2221 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctaggag
     2281 atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     2341 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     2401 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat
     2461 ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac aaaccgtcat
     2521 cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct tcgttattaa
     2581 tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc
     2641 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2701 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2761 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2821 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2881 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt
     2941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3001 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3061 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3121 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3181 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3241 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3301 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3361 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3421 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3481 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3541 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3601 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3661 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3721 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3781 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3841 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3901 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3961 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4021 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4081 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4141 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacatgag
     4201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4381 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     5041 ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa
     5101 tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa
     5161 gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat
     5221 agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg
     5281 gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt
     5341 ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg
     5401 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     5461 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     5521 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     5581 ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     5641 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     5701 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     5761 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     5821 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     5881 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     5941 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     6001 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     6061 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     6121 gaccagttgc gtgatcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     6181 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     6241 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
     6301 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat
     6361 ctgcccttgc ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg
     6421 accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc
     6481 caggactcta gctatagttc tagtggttgg ctacagcttt gtttaaacaa agctggctat
     6541 ggcagggctt gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     6601 ttggggtggg gaaaaggaag aaacgcgggc gtattggtcc caatggggtc tcggtggggt
     6661 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     6721 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct
     6781 ccttccgtgt ttcagttagc ctcccccatc tcccggtacc gcatgctatg catcagctgc
     6841 tagcaccatg gctcgagatc ccgggtgatc aagtcttcgt cgagtgattg taaataaaat
     6901 gtaatttaca gtatagtatt ttaattaata tacaaatgat ttgataataa ttcttattta
     6961 actataatat attgtgttgg gttgaattaa aggtccgtat actagtatcg attcgcgacc
     7021 tactccggaa tattaataga tcatggagat aattaaaatg ataaccatct cgcaaataaa
     7081 taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa tattccggat
     7141 tattcatacc gtcccaccat cgggcgcgga tcctcaatat tggccattag ccatattatt
     7201 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
     7261 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt
     7321 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7381 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     7441 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     7501 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     7561 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     7621 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     7681 tattaccatg
//
