LOCUS       sequence-319250-        6893 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 6893)
  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 6893)
  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
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..206
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     157..177
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     203..227
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          342..474
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     457..476
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    607..1195
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(660..680)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1078..1089)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1328..1462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1365..1384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1419..1438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1491..1656
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1840..2294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1927..1946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2136..2157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2320..2424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2425..2493,2494..3285)
                     /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(2643..2662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3456..4044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3945..3964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4198..4215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4349..4573
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4640..5173)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5362..5367,5368..5384,5385..5390))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    5907..5955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5980..5999
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(6091..6200)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6245..6336
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6261..6278
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     enhancer        join(6516..6893,1..2)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
       61 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      121 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      181 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta
      241 ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct
      301 cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
      361 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
      421 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
      481 actcccagtt caattacagc tcttaaggga tcccggtccg aagcgcgcgg aattcaaagg
      541 cctacgtcga cgagctcact agtcgcggcc gctttcgaat ctagagcctg cagtctcgac
      601 aagcttaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
      661 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
      721 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
      781 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
      841 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
      901 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
      961 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     1021 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     1081 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     1141 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgaa
     1201 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
     1261 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1321 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1441 tgtatcttat catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa
     1501 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     1561 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     1621 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     1681 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     1741 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     1801 gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     1861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2101 ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac
     2161 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2221 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2281 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2341 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2401 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2461 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2521 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2581 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2641 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2701 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2761 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2821 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2881 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2941 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3001 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3061 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3121 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3181 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3241 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3301 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3361 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3421 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3601 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4021 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4081 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4141 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4201 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     4261 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
     4321 taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
     4381 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
     4441 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
     4501 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt
     4561 aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
     4621 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
     4681 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
     4741 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
     4801 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
     4861 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
     4921 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
     4981 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
     5041 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
     5101 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     5161 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
     5221 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
     5281 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgat
     5341 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt
     5401 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag
     5461 gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca
     5521 ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc cttgcttcag
     5581 gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg
     5641 gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat
     5701 agttctagtg gttggctaca gctttgttta aacaaagctg gctatggcag ggcttgccgc
     5761 cccgacgttg gctgcgagcc ctgggccttc acccgaactt gggggttggg gtggggaaaa
     5821 ggaagaaacg cgggcgtatt ggtcccaatg gggtctcggt ggggtatcga cagagtgcca
     5881 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acacccgtgc gttttattct
     5941 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     6001 ttagcctccc ccatctcccg gtaccgcatg ctatgcatca gctgctagca ccatggctcg
     6061 agatcccggg tgatcaagtc ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata
     6121 gtattttaat taatatacaa atgatttgat aataattctt atttaactat aatatattgt
     6181 gttgggttga attaaaggtc cgtatactag tatcgattcg cgacctactc cggaatatta
     6241 atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt attttactgt
     6301 tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc
     6361 accatcgggc gcggatcctc aatattggcc attagccata ttattcattg gttatatagc
     6421 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     6481 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
     6541 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6601 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6661 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6721 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     6781 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6841 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta cca
//
