LOCUS       sequence-438949-        7343 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 7343)
  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 7343)
  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: TH0994-pOCC177-Fus-wt_(1-211)
FEATURES             Location/Qualifiers
     source          1..7343
                     /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             1828..1839
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2108..2128
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2129..2674
                     /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    2810..2944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2847..2866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2901..2920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3017..3389
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3741..3758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3851..4439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3931..3950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4610..5401,5402..5470))
                     /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     5233..5252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5471..5575)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5876..5898
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5890..5907
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5891..5907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5918..5937
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5918..5936
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6547..6671
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6709..7314)
                     /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 gcctcaaacg attataccca acaagcaacc caaagctatg gggcctaccc
     1501 cacccagcca ggacagggct attcccagca gagcagtcag ccctacggac agcagagtta
     1561 cagtggttat agccagtcca cggacacttc aggctatggc cagagcagct attcttctta
     1621 tggccagagc cagaacacag gctatggaac tcagtcaact ccccagggat atggctcgac
     1681 tggcggctat ggcagtagcc agtctagcca atcgtcttac gggcagcagt cctcctaccc
     1741 tggctatggc cagcagccag ctccctcgag cacctcggga agttacggta gcagttctca
     1801 gagcagcagc tatgggcagc cccagagtgg gagctacagc cagcagccta gctatggtgg
     1861 acagcagcaa agctatggac agcagcaaag ctataatccc cctcagggct atggacagca
     1921 gaaccagtac aacagcagct ccggaggtgg aggtggaggt ggaggtggag gtaactatgg
     1981 ccaagatcaa tcctccatga gtagtggtgg tggcagtggt ggcggttatg gcaatcaaga
     2041 ccagagtggt ggaggtggca gcggtggcta tggacagcag ggcgcgccgg gatccagcag
     2101 cggcagagaa aacttgtatt tccagggcat ggacaaagac tgcgaaatga agcgcaccac
     2161 cctggatagc cctctgggca agctggaact gtctgggtgc gaacagggcc tgcaccgtat
     2221 catcttcctg ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg ccccagccgc
     2281 cgtgctgggc ggaccagagc cactgatgca ggccaccgcc tggctcaacg cctactttca
     2341 ccagcctgag gccatcgagg agttccctgt gccagccctg caccacccag tgttccagca
     2401 ggagagcttt acccgccagg tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt
     2461 catcagctac agccacctgg ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa
     2521 aaccgccctg agcggaaatc ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg
     2581 cgacctggac gtggggggct acgagggcgg gctcgccgtg aaagagtggc tgctggccca
     2641 cgagggccac agactgggca agcctgggct gggttaatag aagcttgtcg agaagtacta
     2701 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2761 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2821 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2881 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2941 gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaagc ctgcatccgc
     3001 agatcttaat taataaaaca cgatacattg ttattagtac atttattaag cgctagattc
     3061 tgtgcgttgt tgatttacag acaattgttg tacgtatttt aataattcat taaatttata
     3121 atctttaggg tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa
     3181 tttaaatatt aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg
     3241 ttgtttttcc gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact
     3301 tgccaaatct tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa
     3361 taaaggttcg acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt
     3421 agtgtacaat tgactcgacg taaacacgtt aaataacgct tggacatatt taacatcggg
     3481 cgtgttagct ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc
     3541 ttccgaagac gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc
     3601 cgcgatcaaa tttgtagttg agctttttgg gcctcatggg ccttccgctc actgcccgct
     3661 ttccagtcgg gaaacctgtc gtgccagctg cattaacatg gtcatagctg tttccttgcg
     3721 tattgggcgc tctccgcttc ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag
     3781 cctggggtgc ctaatgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3841 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3901 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3961 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4021 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4081 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4141 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4201 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4261 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4321 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4381 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4441 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4501 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4561 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4621 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4681 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4741 tgatacckcg agarccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4801 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4861 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4921 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4981 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5041 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5101 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5161 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5221 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5281 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5341 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5401 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5461 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5521 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5581 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5641 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5701 atcaaaagaa tagaccgaga tagggttgag tggccgctac agggcgctcc cattcgccat
     5761 tcaggctgcg caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag
     5821 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     5881 tcacgacgtt gtaaaacgac ggccagtgag cgcgacgtaa tacgactcac tatagggcga
     5941 attggcggaa ggccgtcaag gccgtgacgt taaaactatt aagccatcca atcgaccgtt
     6001 agtcgaatca ggaccgctgg tgcgagaagc cgcgaagtat ggcgaatgca tcgtataacg
     6061 tgtggagtcc gctcattaga gcgtcatgtt tagacaagaa agctacatat ttaattgatc
     6121 ccgatgattt tattgataaa ttgaccctaa ctccatacac ggtattctac aatggcgggg
     6181 ttttggtcaa aatttccgga ctgcgattgt acatgctgtt aacggctccg cccactatta
     6241 atgaaattaa aaattccaat tttaaaaaac gcagcaagag aaacatttgt atgaaagaat
     6301 gcgtagaagg aaagaaaaat gtcgtggaca tgctgaacaa caagattaat atgcctccgt
     6361 gtataaaaaa aatattgaac gatttgaaag aaaacaatgt accgcgcggc ggtatgtaca
     6421 ggaagaggtt tatactaaac tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa
     6481 accgatgttt aatcaaggct ctgacgcatt tctacaacca cgactccaag tgtgtgggtg
     6541 aagtcatgca tcttttaatc aaatcccaag atgtgtataa accaccaaac tgccaaaaaa
     6601 tgaaaactgt ggacaagctc tgtccgtttg ctggcaactg caagggtctc aatcctattt
     6661 gtaattattg aataataaaa caattataaa tgctaaattt gttttttatt aacgatacaa
     6721 accaaacgca acaagaacat ttgtagtatt atctataatt gaaaacgcgt agttataatc
     6781 gctgaggtaa tatttaaaat cattttcaaa tgattcacag ttaatttgcg acaatataat
     6841 tttattttca cataaactag acgccttgtc gtcttcttct tcgtattcct tctctttttc
     6901 atttttctcc tcataaaaat taacatagtt attatcgtat ccatatatgt atctatcgta
     6961 tagagtaaat tttttgttgt cataaatata tatgtctttt ttaatggggt gtatagtacc
     7021 gctgcgcata gtttttctgt aatttacaac agtgctattt tctggtagtt cttcggagtg
     7081 tgttgcttta attattaaat ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa
     7141 tatgttgccg gcatagtacg cagcttcttc tagttcaatt acaccatttt ttagcagcac
     7201 cggattaaca taactttcca aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc
     7261 cttttctata ctattgtctg cgagcagttg tttgttgtta aaaataacag ccattgtaat
     7321 gagacgcaca aactaatatc aca
//
