LOCUS       sequence-438950-        8678 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 8678)
  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 8678)
  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: 
            TH1274-054_Jie_EWSR1_pOCC177_pOEM1-N-HIS-MBP-PS-NotI-AscI-TEV-SNAP
FEATURES             Location/Qualifiers
     source          1..8678
                     /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             3443..3463
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3464..4009
                     /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    4145..4279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4182..4201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4236..4255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4352..4724
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(5076..5093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5186..5774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5266..5285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5945..6736,6737..6805))
                     /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     6568..6587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6806..6910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7211..7233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7225..7242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7226..7242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7253..7272
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7253..7271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7882..8006
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(8044..8649)
                     /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 gcgtccacag attactccac ctactcgcaa gcagcagcac agcaaggcta
     1501 ctcagcatac acggcacaac caactcaagg atacgctcag accactcagg cctacggcca
     1561 gcagagctac ggaacttacg gtcagccaac cgacgtgtct tacactcagg ctcagaccac
     1621 tgccacctac ggtcagactg cttacgctac ctcatacgga cagcctccca ccggatacac
     1681 cactccaact gctcctcagg cctactccca gcctgtgcag ggctacggaa ctggtgctta
     1741 cgacaccact accgccaccg tcactaccac tcaggcttca tacgctgccc agtccgccta
     1801 cggaactcag cccgcttacc cagcctacgg tcagcagcct gctgctactg ctcctactcg
     1861 tccccaggac ggaaacaagc ctaccgagac ttcacagccc cagtccagca ccggtggcta
     1921 caaccagccc tccctgggct acggacagtc aaactactcc tacccacagg tgcctggttc
     1981 ctaccccatg cagccagtca ccgccccacc tagctaccct cctacttctt actcttcaac
     2041 ccagccaact tcatacgacc agtccagcta ctcccagcag aacacctacg gacagccttc
     2101 ttcatacggt cagcagtcca gctacggcca gcagtcttca tacggacagc agcctcccac
     2161 cagctaccca cctcagactg gatcatactc ccaggctccc agccagtact ctcagcagtc
     2221 cagctcttac ggtcagcagt catccttccg ccaggaccac ccaagctcta tgggcgtgta
     2281 cggacaggag agcggaggtt tctctggtcc tggcgaaaac agaagcatgt ctggacctga
     2341 caaccgcgga cgtggtaggg gcggattcga ccgcggtggc atgtctcgtg gaggtagggg
     2401 cggaggtcgt ggcggaatgg gttctgctgg cgagaggggt ggcttcaaca agcctggagg
     2461 tcccatggac gaaggtccag acctggacct gggtcctcct gtcgaccctg acgaggactc
     2521 agacaactcc gctatctacg tgcagggtct gaacgactcc gtcactctgg acgacctggc
     2581 cgacttcttc aagcagtgcg gcgtggtcaa gatgaacaag cgtaccggac agcccatgat
     2641 ccacatctac ctggacaagg aaaccggaaa gccaaagggt gacgctactg tgagctacga
     2701 ggaccctccc actgctaagg ctgctgtcga atggttcgac ggcaaggact tccagggctc
     2761 aaagctgaag gtgtccctgg ctagaaagaa gccacctatg aactctatgc gcggcggact
     2821 gcccccaaga gaaggaaggg gaatgcctcc cccactgagg ggtggcccag gaggtcctgg
     2881 cggacccggt ggcccaatgg gaaggatggg aggtagaggc ggagaccgcg gtggcttccc
     2941 tccccgtgga ccaaggggta gcagaggcaa cccatctgga ggtggcaacg tgcaacacag
     3001 ggctggtgac tggcagtgcc caaacccagg ttgcggcaac cagaacttcg cttggaggac
     3061 cgagtgcaac cagtgcaagg cccctaagcc cgaaggtttc ctgccacctc ccttcccacc
     3121 tcccggaggt gacagaggcc gcggcggacc tggtggcatg cgcggaggtc gtggcggact
     3181 gatggacaga ggtggccccg gaggtatgtt caggggcgga agaggtggcg accgtggagg
     3241 tttccgcggc ggacgtggaa tggacagggg tggcttcgga ggtggccgcc gtggaggtcc
     3301 aggcggacca cctggtcctc tgatggaaca gatgggtggc aggagaggag gtcgcggtgg
     3361 accaggaaaa atggataaag gagaacacag acaggagcgt cgtgacagac cctacggcgc
     3421 gccgggatcc agcagcggca gagaaaactt gtatttccag ggcatggaca aagactgcga
     3481 aatgaagcgc accaccctgg atagccctct gggcaagctg gaactgtctg ggtgcgaaca
     3541 gggcctgcac cgtatcatct tcctgggcaa aggaacatct gccgccgacg ccgtggaagt
     3601 gcctgcccca gccgccgtgc tgggcggacc agagccactg atgcaggcca ccgcctggct
     3661 caacgcctac tttcaccagc ctgaggccat cgaggagttc cctgtgccag ccctgcacca
     3721 cccagtgttc cagcaggaga gctttacccg ccaggtgctg tggaaactgc tgaaagtggt
     3781 gaagttcgga gaggtcatca gctacagcca cctggccgcc ctggccggca atcccgccgc
     3841 caccgccgcc gtgaaaaccg ccctgagcgg aaatcccgtg cccattctga tcccctgcca
     3901 ccgggtggtg cagggcgacc tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga
     3961 gtggctgctg gcccacgagg gccacagact gggcaagcct gggctgggtt aatagaagct
     4021 tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4081 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4141 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4201 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4261 atcttatcat gtctggatct gatcactgct tgagcctagg agatccgaac cagataagtg
     4321 aaagcctgca tccgcagatc ttaattaata aaacacgata cattgttatt agtacattta
     4381 ttaagcgcta gattctgtgc gttgttgatt tacagacaat tgttgtacgt attttaataa
     4441 ttcattaaat ttataatctt tagggtggta tgttagagcg aaaatcaaat gattttcagc
     4501 gtctttatat ctgaatttaa atattaaatc ctcaatagat ttgtaaaata ggtttcgatt
     4561 agtttcaaac aagggttgtt tttccgaacc gatggctgga ctatctaatg gattttcgct
     4621 caacgccaca aaacttgcca aatcttgtag cagcaatcta gctttgtcga tattcgtttg
     4681 tgttttgttt tgtaataaag gttcgacgtc gttcaaaata ttatgcgctt ttgtatttct
     4741 ttcatcactg tcgttagtgt acaattgact cgacgtaaac acgttaaata acgcttggac
     4801 atatttaaca tcgggcgtgt tagctttatt aggccgatta tcgtcgtcgt cccaaccctc
     4861 gtcgttagaa gttgcttccg aagacgattt tgccatagcc acacgacgcc tattaattgt
     4921 gtcggctaac acgtccgcga tcaaatttgt agttgagctt tttgggcctc atgggccttc
     4981 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat
     5041 agctgtttcc ttgcgtattg ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg
     5101 gtcgttcggg taaagcctgg ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc
     5161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5881 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5941 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6001 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6061 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa
     6121 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6181 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6241 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6301 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6361 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6421 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6481 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6541 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6601 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6661 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6721 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6781 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6841 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6901 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     6961 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7021 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtggcc gctacagggc
     7081 gctcccattc gccattcagg ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct
     7141 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7201 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcga cgtaatacga
     7261 ctcactatag ggcgaattgg cggaaggccg tcaaggccgt gacgttaaaa ctattaagcc
     7321 atccaatcga ccgttagtcg aatcaggacc gctggtgcga gaagccgcga agtatggcga
     7381 atgcatcgta taacgtgtgg agtccgctca ttagagcgtc atgtttagac aagaaagcta
     7441 catatttaat tgatcccgat gattttattg ataaattgac cctaactcca tacacggtat
     7501 tctacaatgg cggggttttg gtcaaaattt ccggactgcg attgtacatg ctgttaacgg
     7561 ctccgcccac tattaatgaa attaaaaatt ccaattttaa aaaacgcagc aagagaaaca
     7621 tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt ggacatgctg aacaacaaga
     7681 ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt gaaagaaaac aatgtaccgc
     7741 gcggcggtat gtacaggaag aggtttatac taaactgtta cattgcaaac gtggtttcgt
     7801 gtgccaagtg tgaaaaccga tgtttaatca aggctctgac gcatttctac aaccacgact
     7861 ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc ccaagatgtg tataaaccac
     7921 caaactgcca aaaaatgaaa actgtggaca agctctgtcc gtttgctggc aactgcaagg
     7981 gtctcaatcc tatttgtaat tattgaataa taaaacaatt ataaatgcta aatttgtttt
     8041 ttattaacga tacaaaccaa acgcaacaag aacatttgta gtattatcta taattgaaaa
     8101 cgcgtagtta taatcgctga ggtaatattt aaaatcattt tcaaatgatt cacagttaat
     8161 ttgcgacaat ataattttat tttcacataa actagacgcc ttgtcgtctt cttcttcgta
     8221 ttccttctct ttttcatttt tctcctcata aaaattaaca tagttattat cgtatccata
     8281 tatgtatcta tcgtatagag taaatttttt gttgtcataa atatatatgt cttttttaat
     8341 ggggtgtata gtaccgctgc gcatagtttt tctgtaattt acaacagtgc tattttctgg
     8401 tagttcttcg gagtgtgttg ctttaattat taaatttata taatcaatga atttgggatc
     8461 gtcggttttg tacaatatgt tgccggcata gtacgcagct tcttctagtt caattacacc
     8521 attttttagc agcaccggat taacataact ttccaaaatg ttgtacgaac cgttaaacaa
     8581 aaacagttca cctccctttt ctatactatt gtctgcgagc agttgtttgt tgttaaaaat
     8641 aacagccatt gtaatgagac gcacaaacta atatcaca
//
