LOCUS       sequence-466606-        8294 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 8294)
  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 8294)
  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: TH1150-pOCC177-FUS(Nhe)-PLD_YtoF_RGG_RtoK
FEATURES             Location/Qualifiers
     source          1..8294
                     /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             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..1360
                     /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     1334..1357
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1415..1438
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3059..3079
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3080..3625
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    3761..3895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3798..3817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3852..3871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3968..4340
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4692..4709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4802..5390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4882..4901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5561..6352,6353..6421))
                     /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     6184..6203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6422..6526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6827..6849
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6841..6858
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6842..6858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6869..6888
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6869..6887
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7498..7622
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7660..8265)
                     /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 ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagaa tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg
      301 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
      361 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
      421 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
      481 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
      541 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
      601 atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc
      661 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
      721 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
      781 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
      841 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
      901 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
      961 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     1021 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     1081 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     1141 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggtgaa
     1201 agatccgcgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     1261 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     1321 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct cgaacaacaa
     1381 caacaataac aataacaaca acagcagcgg cagactggaa gttctgttcc aggggccggc
     1441 ggccgcaatg gcgagcaacg acttcactca acaggcaact caatcgttcg gtgcatttcc
     1501 gacacagcca ggccagggat tctctcagca gtcttcgcag ccgtttggcc agcagtcttt
     1561 ctccggattc tcccaaagca cagatactag tggttttggc caatcctcct ttagttcatt
     1621 cggtcaaagc cagaacactg gtttcggaac acagagcacc ccgcaaggat ttggctcgac
     1681 gggtggcttc ggttcgagtc agagcagtca atcgtctttc ggacaacagt ctagctttcc
     1741 tggtttcgga caacaacctg caccaagcag cacctcggga agtttcggca gttcctccca
     1801 gtcctcgtca ttcggccaac cgcagtcggg ctcatttagt caacaaccga gttttggcgg
     1861 acagcaacag tcatttggac agcaacaatc tttcaaccct ccacagggtt ttggacagca
     1921 aaaccagttt aatagttcta gcggtggtgg cggaggtggc ggtggtggcg gaaacttcgg
     1981 ccaggaccaa tcaagcatgt caagcggcgg aggctccggc ggaggctttg gaaaccaaga
     2041 ccaaagcggc ggtggcggtt cgggaggttt tggtcagcag gctagcgata agggaggaaa
     2101 gggtaagggc ggctcgggtg gcggaggcgg cggtggaggc ggaggctaca atcgctcttc
     2161 cggaggttat gagccgaagg gcaagggtgg cggaaaaggt ggaaagggag gaatgggcgg
     2221 tagtgacaag ggtggattca acaaattcgg tggacctcgc gatcagggta gtagacatga
     2281 tagtgagcaa gacaattcag ataataatac aatttttgtg cagggcttgg gagagaacgt
     2341 taccattgag tcggttgcgg actactttaa gcagatcggt ataattaaga ccaacaaaaa
     2401 gactggccag cccatgataa atctgtacac tgaccgcgag acgggcaagc tgaaaggcga
     2461 agctacagtg tctttcgacg atcctccatc cgccaaagcg gcgatcgact ggtttgacgg
     2521 aaaagagttc agcggtaatc cgatcaaggt ctctttcgca acgcgtaggg cagatttcaa
     2581 caagggaggc ggtaatggta aaggcggcaa aggtaaggga ggcccgatgg gcaaaggcgg
     2641 atacggcgga ggcggcagcg gcggaggagg caaaggtgga tttccgagcg gcggaggcgg
     2701 aggtggtggc caacagcgtg caggagattg gaagtgtccc aacccgacat gcgagaatat
     2761 gaattttagt tggcgtaatg agtgtaatca atgtaaagcc ccaaaacctg atggtcctgg
     2821 cggcggccct ggaggttccc atatgggagg taactacggt gatgatagaa agggcggcaa
     2881 aggaggatac gacaagggag gttataaagg aaaaggaggc gataagggag gcttccgagg
     2941 cggccggggt ggtggggaca gaggtggctt tggccctggc aagatggatt ccaggggtga
     3001 gcacagacag gatcgcaggg agaggccgta tggcgcgccg ggatccagca gcggcagaga
     3061 aaacttgtat ttccagggca tggacaaaga ctgcgaaatg aagcgcacca ccctggatag
     3121 ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct
     3181 gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg
     3241 cggaccagag ccactgatgc aggccaccgc ctggctcaac gcctactttc accagcctga
     3301 ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc aggagagctt
     3361 tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttcggagagg tcatcagcta
     3421 cagccacctg gccgccctgg ccggcaatcc cgccgccacc gccgccgtga aaaccgccct
     3481 gagcggaaat cccgtgccca ttctgatccc ctgccaccgg gtggtgcagg gcgacctgga
     3541 cgtggggggc tacgagggcg ggctcgccgt gaaagagtgg ctgctggccc acgagggcca
     3601 cagactgggc aagcctgggc tgggttaata gaagcttgtc gagaagtact agaggatcat
     3661 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3721 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3781 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3841 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     3901 actgcttgag cctaggagat ccgaaccaga taagtgaaag cctgcatccg cagatcttaa
     3961 ttaataaaac acgatacatt gttattagta catttattaa gcgctagatt ctgtgcgttg
     4021 ttgatttaca gacaattgtt gtacgtattt taataattca ttaaatttat aatctttagg
     4081 gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga atttaaatat
     4141 taaatcctca atagatttgt aaaataggtt tcgattagtt tcaaacaagg gttgtttttc
     4201 cgaaccgatg gctggactat ctaatggatt ttcgctcaac gccacaaaac ttgccaaatc
     4261 ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta ataaaggttc
     4321 gacgtcgttc aaaatattat gcgcttttgt atttctttca tcactgtcgt tagtgtacaa
     4381 ttgactcgac gtaaacacgt taaataacgc ttggacatat ttaacatcgg gcgtgttagc
     4441 tttattaggc cgattatcgt cgtcgtccca accctcgtcg ttagaagttg cttccgaaga
     4501 cgattttgcc atagccacac gacgcctatt aattgtgtcg gctaacacgt ccgcgatcaa
     4561 atttgtagtt gagctttttg ggcctcatgg gccttccgct cactgcccgc tttccagtcg
     4621 ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc gtattgggcg
     4681 ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa gcctggggtg
     4741 cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5221 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5641 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5701 gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5761 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5821 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5881 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5941 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6001 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6061 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6121 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6181 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6241 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6301 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6361 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6421 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6541 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6601 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6661 atagaccgag atagggttga gtggccgcta cagggcgctc ccattcgcca ttcaggctgc
     6721 gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6781 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6841 tgtaaaacga cggccagtga gcgcgacgta atacgactca ctatagggcg aattggcgga
     6901 aggccgtcaa ggccgtgacg ttaaaactat taagccatcc aatcgaccgt tagtcgaatc
     6961 aggaccgctg gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac gtgtggagtc
     7021 cgctcattag agcgtcatgt ttagacaaga aagctacata tttaattgat cccgatgatt
     7081 ttattgataa attgacccta actccataca cggtattcta caatggcggg gttttggtca
     7141 aaatttccgg actgcgattg tacatgctgt taacggctcc gcccactatt aatgaaatta
     7201 aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag
     7261 gaaagaaaaa tgtcgtggac atgctgaaca acaagattaa tatgcctccg tgtataaaaa
     7321 aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt
     7381 ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt
     7441 taatcaaggc tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc
     7501 atcttttaat caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg
     7561 tggacaagct ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt
     7621 gaataataaa acaattataa atgctaaatt tgttttttat taacgataca aaccaaacgc
     7681 aacaagaaca tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta
     7741 atatttaaaa tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc
     7801 acataaacta gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc
     7861 ctcataaaaa ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa
     7921 ttttttgttg tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat
     7981 agtttttctg taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt
     8041 aattattaaa tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc
     8101 ggcatagtac gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac
     8161 ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat
     8221 actattgtct gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac
     8281 aaactaatat caca
//
