LOCUS       sequence-438938-        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: TH1203-pOCC177_pOEM1-N-HIS-MBP-PS-TAF15_opt-TE...
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             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             3242..3262
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3263..3808
                     /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    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 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 agcgatagtg gaagttacgg acagtcggga ggcgagcagc aaagctacag
     1501 cacctacgga aaccctggaa gtcaaggata cggacaggct tcccagagct actctggcta
     1561 cggacagacc actgactcca gctacggaca gaactactct ggttactctt catacggcca
     1621 gtcatactcc cagagctacg gtggctacga gaaccagaag cagtccagct actcacagca
     1681 gccatacaac aaccagggcc agcagcagaa catggaatct tcaggttctc agggaggtcg
     1741 cgccccttca tacgaccagc ccgactacgg tcagcaggac tcctacgacc agcagagcgg
     1801 ttacgaccag caccagggct cttacgacga gcagtcaaac tacgaccagc agcacgactc
     1861 ttactcacag aaccagcagt cttaccactc acagcgcgag aactactctc accacactca
     1921 agacgacagg agggacgtga gcaggtacgg tgaagacaac aggggttacg gcggaagcca
     1981 gggtggcgga cgcggccgtg gtggctacga caaggacggt agaggcccca tgactggttc
     2041 cagcggaggt gaccgcggcg gattcaagaa cttcggtggc caccgcgact acggtccacg
     2101 tactgacgct gactccgaga gcgacaactc cgacaacaac accatcttcg tccagggact
     2161 gggagagggc gtgagcactg accaggtcgg cgaattcttc aagcagatcg gtatcatcaa
     2221 gaccaacaag aagactggca agccaatgat caacctgtac accgacaagg acactggcaa
     2281 gcctaagggc gaagctaccg tgtccttcga cgaccctccc agcgccaagg ctgccatcga
     2341 ctggttcgac ggcaaggagt tccacggaaa catcatcaag gtctctttcg ctactaggag
     2401 acctgaattc atgcgtggag gtggctcagg aggtggccgc cgtggaaggg gaggttacag
     2461 gggcagaggc ggattccagg gtcgtggtgg cgacccaaag tctggcgact gggtgtgccc
     2521 aaacccttca tgcggtaaca tgaacttcgc caggagaaac tcctgcaacc agtgcaacga
     2581 gcccaggcca gaagactcca gacctagcgg aggtgacttc cgcggacgtg gttacggcgg
     2641 agaaaggggc tacaggggaa gaggtggcag gggaggtgac agaggcggat acggtggcga
     2701 cagatctgga ggtggctacg gaggtgaccg ctcttcaggc ggaggttact ctggtgaccg
     2761 ttcaggcgga ggttacggcg gagaccgttc cggtggcgga tatggtggcg accgcggagg
     2821 tggctatgga ggtgaccgtg gcggaggtta tggcggagac aggggtggcg gatacggtgg
     2881 cgacagggga ggttatggcg gagacagagg tggcggatac ggtggcgacc gcggaggtta
     2941 tggcggagac cgtggtggct atggaggtga caggggcgga tatggtggcg acagaggagg
     3001 ttatggcgga gacaggtctc gtggtggcta cggaggtgac cgcggcggag gttctggtta
     3061 cggcggagac agaagcggtg gctatggagg tgacaggagc ggcggaggtt atggcggaga
     3121 ccgtggtggc ggatacggtg gcgacagggg aggttacgga ggaaagatgg gaggcagaaa
     3181 cgactaccgc aacgaccaaa ggaacagacc ctacggcgcg ccgggatcca gcagcggcag
     3241 agaaaacttg tatttccagg gcatggacaa agactgcgaa atgaagcgca ccaccctgga
     3301 tagccctctg ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc gtatcatctt
     3361 cctgggcaaa ggaacatctg ccgccgacgc cgtggaagtg cctgccccag ccgccgtgct
     3421 gggcggacca gagccactga tgcaggccac cgcctggctc aacgcctact ttcaccagcc
     3481 tgaggccatc gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc agcaggagag
     3541 ctttacccgc caggtgctgt ggaaactgct gaaagtggtg aagttcggag aggtcatcag
     3601 ctacagccac ctggccgccc tggccggcaa tcccgccgcc accgccgccg tgaaaaccgc
     3661 cctgagcgga aatcccgtgc ccattctgat cccctgccac cgggtggtgc agggcgacct
     3721 ggacgtgggg ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg cccacgaggg
     3781 ccacagactg ggcaagcctg ggctgggtta 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
//
