LOCUS       sequence-438941-        8477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector for baculovirus production, using FlashBac bacmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8477)
  AUTHORS   Wang J, Choi JM, Holehouse AS, Lee HO, Zhang X, Jahnel M, Maharana 
            S, Lemaitre R, Pozniakovsky A, Drechsel D, Poser I, Pappu RV, 
            Alberti S, Hyman AA
  TITLE     A Molecular Grammar Governing the Driving Forces for Phase 
            Separation of Prion-like RNA Binding Proteins.
  JOURNAL   Cell. 2018 Jul 26;174(3):688-699.e16. doi: 
            10.1016/j.cell.2018.06.006. Epub 2018 Jun 28.
  PUBMED    29961577
REFERENCE   2  (bases 1 to 8477)
  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: 
            "10.1016/j.cell.2018.06.006"; journalName: "Cell"; date: 
            "2018-07-26- 26"; volume: "174"; issue: "3"; pages: "688-699.e16"
COMMENT     Sequence Label: TH1204-pOCC120-FUS_opt(NheI)
FEATURES             Location/Qualifiers
     source          1..8477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..170
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     95..112
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             224..1324
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1298..1321
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1379..1402
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1792..1803
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             join(3035..3037,3038..3040,3041..3751)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3080..3101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3341..3360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3688..3709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3752..3775
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3785..3802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3944..4078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3981..4000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4035..4054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4151..4523
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4875..4892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4985..5573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5065..5084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5744..6535,6536..6604))
                     /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     6367..6386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6605..6709)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7010..7032
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7024..7041
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7025..7041
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7052..7071
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7052..7070
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7681..7805
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7843..8448)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 agctgccacc attttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
       61 actccggaat attaatagat catggagata attaaaatga taaccatctc gcaaataaat
      121 aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt
      181 attcataccg tcccaccatc gggcgcggat cgccaccatg ggcatgaaaa tcgaagaagg
      241 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      301 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      361 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      421 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      481 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      541 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      601 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      661 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      721 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      781 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      841 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      901 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      961 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
     1021 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1081 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1141 gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca
     1201 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1261 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1321 gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca gcggcagact
     1381 ggaagttctg ttccaggggc cggcggccgc aatggcctca aacgattata cccaacaagc
     1441 aacccaaagc tatggggcct accccaccca gccaggacag ggctattccc agcagagcag
     1501 tcagccctac ggacagcaga gttacagtgg ttatagccag tccacggaca cttcaggcta
     1561 tggccagagc agctattctt cttatggcca gagccagaac acaggctatg gaactcagtc
     1621 aactccccag ggatatggct cgactggcgg ctatggcagt agccagtcta gccaatcgtc
     1681 ttacgggcag cagtcctcct accctggcta tggccagcag ccagctccct cgagcacctc
     1741 gggaagttac ggtagcagtt ctcagagcag cagctatggg cagccccaga gtgggagcta
     1801 cagccagcag cctagctatg gtggacagca gcaaagctat ggacagcagc aaagctataa
     1861 tccccctcag ggctatggac agcagaacca gtacaacagc agctccggag gtggaggtgg
     1921 aggtggaggt ggaggtaact atggccaaga tcaatcctcc atgagtagtg gtggtggcag
     1981 tggtggcggt tatggcaatc aagaccagag tggtggaggt ggcagcggtg gctatggaca
     2041 gcaggctagc gaccgtggag gacgaggcag gggtggatcc ggtggcggcg gcggcggcgg
     2101 cggtggtggt tataaccgca gcagtggtgg ctatgaaccc agaggtcgtg gaggtggccg
     2161 tggaggcaga ggtggcatgg gcggaagtga ccgtggtggc ttcaataaat ttggtggccc
     2221 gcgggaccaa ggatcacgtc atgactccga acaggataat tcagacaaca acaccatctt
     2281 tgtacaaggc ctgggtgaga atgttacaat tgagtctgtg gctgattact tcaagcagat
     2341 tggtattatt aagacaaaca agaaaacggg acagcccatg attaatctct acacagacag
     2401 ggaaactggc aagctgaagg gagaggcaac ggtctctttt gatgacccac cttcagctaa
     2461 agcagctatt gactggtttg atggtaaaga gttctctgga aatcctatca aggtctcatt
     2521 tgctacgcgt cgggcagact ttaatcgggg tggtggcaat ggtcgtggag gccgagggcg
     2581 aggaggacca atgggccgtg gaggctatgg aggtggtggc agtggtggtg gtggccgagg
     2641 aggatttccc agtggaggtg gtggcggtgg aggacagcag cgagccggcg actggaagtg
     2701 tcctaatccc acctgtgaga atatgaactt ctcttggagg aatgaatgca accagtgtaa
     2761 ggcccctaaa ccagatggcc cgggaggggg accaggtggc tctcacatgg ggggtaacta
     2821 cggggatgat cgtcgtggtg gcagaggagg ctatgatcga ggcggctacc ggggacgagg
     2881 cggcgatcgc ggaggcttcc gaggcggccg gggtggtggg gacagaggtg gctttggccc
     2941 tggcaagatg gattccaggg gtgagcacag acaggatcgc agggagaggc cgtatggcgc
     3001 gccgggatcc gctggctccg ctgctggttc tggcatggtg agcaagggcg aggagctgtt
     3061 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3121 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3181 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3241 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3301 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3361 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3421 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3481 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3541 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3601 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaaactgag
     3661 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3721 gatcactctc ggcatggacg agctgtacaa gctggaagtt ctgttccagg ggccgggcag
     3781 cagccatcac catcaccatc acagtggata atagaagctt gtcgagaagt actagaggat
     3841 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3901 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3961 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4021 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg
     4081 atcactgctt gagcctagga gatccgaacc agataagtga aagcctgcat ccgcagatct
     4141 taattaataa aacacgatac attgttatta gtacatttat taagcgctag attctgtgcg
     4201 ttgttgattt acagacaatt gttgtacgta ttttaataat tcattaaatt tataatcttt
     4261 agggtggtat gttagagcga aaatcaaatg attttcagcg tctttatatc tgaatttaaa
     4321 tattaaatcc tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca agggttgttt
     4381 ttccgaaccg atggctggac tatctaatgg attttcgctc aacgccacaa aacttgccaa
     4441 atcttgtagc agcaatctag ctttgtcgat attcgtttgt gttttgtttt gtaataaagg
     4501 ttcgacgtcg ttcaaaatat tatgcgcttt tgtatttctt tcatcactgt cgttagtgta
     4561 caattgactc gacgtaaaca cgttaaataa cgcttggaca tatttaacat cgggcgtgtt
     4621 agctttatta ggccgattat cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga
     4681 agacgatttt gccatagcca cacgacgcct attaattgtg tcggctaaca cgtccgcgat
     4741 caaatttgta gttgagcttt ttgggcctca tgggccttcc gctcactgcc cgctttccag
     4801 tcgggaaacc tgtcgtgcca gctgcattaa catggtcata gctgtttcct tgcgtattgg
     4861 gcgctctccg cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg
     4921 gtgcctaatg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5401 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5881 cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6721 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     6781 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6841 agaatagacc gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc
     6901 tgcgcaactg ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg
     6961 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     7021 cgttgtaaaa cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattggc
     7081 ggaaggccgt caaggccgtg acgttaaaac tattaagcca tccaatcgac cgttagtcga
     7141 atcaggaccg ctggtgcgag aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga
     7201 gtccgctcat tagagcgtca tgtttagaca agaaagctac atatttaatt gatcccgatg
     7261 attttattga taaattgacc ctaactccat acacggtatt ctacaatggc ggggttttgg
     7321 tcaaaatttc cggactgcga ttgtacatgc tgttaacggc tccgcccact attaatgaaa
     7381 ttaaaaattc caattttaaa aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag
     7441 aaggaaagaa aaatgtcgtg gacatgctga acaacaagat taatatgcct ccgtgtataa
     7501 aaaaaatatt gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga
     7561 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
     7621 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
     7681 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
     7741 ctgtggacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
     7801 attgaataat aaaacaatta taaatgctaa atttgttttt tattaacgat acaaaccaaa
     7861 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
     7921 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
     7981 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
     8041 ctcctcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
     8101 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
     8161 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
     8221 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
     8281 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
     8341 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
     8401 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
     8461 cacaaactaa tatcaca
//
