LOCUS       sequence-319256-        7696 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 7696)
  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 7696)
  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 myc
FEATURES             Location/Qualifiers
     source          1..7696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      386..974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     875..894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1128..1145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1279..1503
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1570..2103)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2292..2297,2298..2314,2315..2320))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    2837..2885
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2910..2929
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(3021..3130)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        3175..3266
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3191..3208
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     enhancer        3446..3825
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3826..4029
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3980..4000
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4026..4050
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4165..4297
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4280..4299
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4409..4432
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4445..4447,4448..5158)
                     /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(4487..4508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4748..4767)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5095..5116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5174..5203
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5233..5821
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5286..5306)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5704..5715)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5954..6088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5991..6010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6045..6064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6117..6282
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6466..6920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6553..6572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6762..6783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6946..7050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7051..7119,7120..7696,1..215)
                     /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(7269..7288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
       61 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      121 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      181 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
      241 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
      301 ttgataatct catgaccaaa atcccttaac atgagttttc gttccactga gcgtcagacc
      361 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      421 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      481 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      541 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      601 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      661 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      721 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      781 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      841 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      901 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      961 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1021 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1081 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1141 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1201 cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag taataaatgg
     1261 atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca aaatagatct
     1321 aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa tcagtccagt
     1381 tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct tcattttctg
     1441 aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt aaagactata
     1501 ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg
     1561 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
     1621 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
     1681 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
     1741 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
     1801 ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     1861 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     1921 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     1981 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     2041 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     2101 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     2161 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     2221 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgat cgcatacgct
     2281 acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc
     2341 gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt
     2401 cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct
     2461 tgctgttctt ctacggcaag gtgctgtgca cggatctgcc cttgcttcag gagatcggta
     2521 gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc
     2581 tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat agttctagtg
     2641 gttggctaca gctttgttta aacaaagctg gctatggcag ggcttgccgc cccgacgttg
     2701 gctgcgagcc ctgggccttc acccgaactt gggggttggg gtggggaaaa ggaagaaacg
     2761 cgggcgtatt ggtcccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     2821 cgaaccccgc gtttatgaac aaacgaccca acacccgtgc gttttattct gtctttttat
     2881 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc
     2941 ccatctcccg gtaccgcatg ctatgcatca gctgctagca ccatggctcg agatcccggg
     3001 tgatcaagtc ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata gtattttaat
     3061 taatatacaa atgatttgat aataattctt atttaactat aatatattgt gttgggttga
     3121 attaaaggtc cgtatactag tatcgattcg cgacctactc cggaatatta atagatcatg
     3181 gagataatta aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca
     3241 gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc
     3301 gcggatcctc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     3361 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     3421 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
     3481 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3541 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3601 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3661 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
     3721 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
     3781 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     3841 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     3901 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3961 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4021 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
     4081 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcaga
     4141 agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac
     4201 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
     4261 ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac
     4321 agctcttaag ggatcccggt ccgaagcgcg cggaattcaa aggcctacgt cgacgagctc
     4381 actagtcgcg gccgctttcg aatctagact ggaagttctg ttccagggtc ctgcagccgc
     4441 cgcggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4501 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4561 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4621 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     4681 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4741 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4801 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4861 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     4921 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     4981 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     5041 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     5101 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     5161 cggaggttca ggagaacaaa aactcatctc agaagaggat ctgtgactag agcctgcagt
     5221 ctcgacaagc ttaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5281 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5341 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5401 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5461 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5521 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5581 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     5641 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5701 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5761 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5821 cctgaagctt gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt
     5881 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5941 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6061 catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga gatccgaacc
     6121 agataagtga aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg
     6181 acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc
     6241 atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca tttttcttcc
     6301 tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca tcttcggcta
     6361 ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa
     6421 ttgactgaat atcaacgctt atttgcagcc tgaatggcga atgggacgcg ccctgtagcg
     6481 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6541 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6601 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6661 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6721 cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct tgttccaaac
     6781 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6841 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6901 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6961 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     7021 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     7081 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     7141 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     7201 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     7261 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     7321 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7381 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7441 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7501 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7561 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7621 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7681 gcggataaag ttgcag
//
