LOCUS       sequence-438937-        8015 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 8015)
  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 8015)
  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: 
            TH1202-pOCC175_pOEM1-N-HIS-MBP-PS-mGFP-TEV-HNRNPA3_opt
FEATURES             Location/Qualifiers
     source          1..8015
                     /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             join(1439..1441,1442..1444,1445..2155)
                     /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(1484..1505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1745..1764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2092..2113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2174..2194
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3482..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3519..3538)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3573..3592
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3689..4061
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4413..4430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4523..5111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4603..4622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5282..6073,6074..6142))
                     /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     5905..5924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6143..6247)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6548..6570
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6562..6579
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6563..6579
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6590..6609
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6590..6608
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7219..7343
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7381..7986)
                     /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 aggggccgat
     1441 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1501 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1561 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1621 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1681 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1741 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1801 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1861 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1921 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1981 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2041 cctgagcacc cagtccaaac tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2101 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggatc
     2161 cagcagcggc agagaaaact tgtatttcca gggcgcggcc gcaatggaag tgaaaccccc
     2221 accaggaaga ccacagcccg acagcggtag aagaagaaga agaagaggag aggagggaca
     2281 cgacccgaaa gagcccgaac agctgaggaa gctgttcatc ggtggcctgt ccttcgaaac
     2341 cactgacgac agcctgcgcg agcacttcga aaagtggggt accctgactg actgcgtggt
     2401 catgcgtgac ccacagacca agcgctcccg tggattcggt ttcgtgactt acagctgcgt
     2461 ggaggaagtc gacgctgcca tgtgcgctcg ccctcacaag gtggacggaa gggtggtgga
     2521 accaaagagg gctgtgtccc gtgaggacag cgtcaagcct ggtgcccacc tgaccgtgaa
     2581 gaagatcttc gtcggaggta tcaaggaaga cactgaggaa tacaacctgc gcgactactt
     2641 cgagaagtac ggcaagatcg agaccatcga agtgatggag gacaggcagt ccggcaagaa
     2701 gcgcggcttc gccttcgtga ccttcgacga ccacgacact gtcgacaaga tcgtggtcca
     2761 gaagtaccac actatcaacg gtcacaactg cgaagtcaag aaggctctgt ctaagcagga
     2821 gatgcaatct gctggaagcc agaggggtag aggcggaggt agcggcaact tcatgggacg
     2881 tggcggaaac ttcggtggcg gaggtggcaa cttcggtagg ggaggtaact tcggcggaag
     2941 aggtggctac ggaggtggcg gaggtggctc taggggatca tacggaggtg gcgacggagg
     3001 ttacaacggt ttcggcggag acggtggcaa ctacggaggt ggcccaggct actccagcag
     3061 aggaggttac ggcggaggtg gccctggcta cggaaaccag ggaggtggct atggaggtgg
     3121 cggaggttac gacggttaca acgagggcgg aaattttggt ggcggaaact acggtggcgg
     3181 aggtaactac aacgacttcg gtaactactc tggccagcag cagtcaaact acggccccat
     3241 gaagggcgga tctttcggtg gccgttcttc aggatcacca tacggtggtg gttacggcag
     3301 tggtggtggt agcggtggtt acggcagcag aaggttcggc gcgccgtaat agaagcttgt
     3361 cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3421 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3481 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3601 ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag ataagtgaaa
     3661 gcctgcatcc gcagatctta attaataaaa cacgatacat tgttattagt acatttatta
     3721 agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt ttaataattc
     3781 attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc
     3841 tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt ttcgattagt
     3901 ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat tttcgctcaa
     3961 cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat tcgtttgtgt
     4021 tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg tatttctttc
     4081 atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaataacg cttggacata
     4141 tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc
     4201 gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat taattgtgtc
     4261 ggctaacacg tccgcgatca aatttgtagt tgagcttttt gggcctcatg ggccttccgc
     4321 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc
     4381 tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc
     4441 gttcgggtaa agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta
     4501 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4561 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4621 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4681 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4741 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4801 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4861 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4921 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4981 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5041 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5101 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5161 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5221 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5281 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5341 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5401 ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa
     5461 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5521 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5581 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5641 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5701 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5761 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5821 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5881 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5941 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6001 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6061 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6121 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6181 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6241 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6301 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6361 aatcccttat aaatcaaaag aatagaccga gatagggttg agtggccgct acagggcgct
     6421 cccattcgcc attcaggctg cgcaactgtt gggaagggcg tttcggtgcg ggcctcttcg
     6481 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6541 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgacgt aatacgactc
     6601 actatagggc gaattggcgg aaggccgtca aggccgtgac gttaaaacta ttaagccatc
     6661 caatcgaccg ttagtcgaat caggaccgct ggtgcgagaa gccgcgaagt atggcgaatg
     6721 catcgtataa cgtgtggagt ccgctcatta gagcgtcatg tttagacaag aaagctacat
     6781 atttaattga tcccgatgat tttattgata aattgaccct aactccatac acggtattct
     6841 acaatggcgg ggttttggtc aaaatttccg gactgcgatt gtacatgctg ttaacggctc
     6901 cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag agaaacattt
     6961 gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtgga catgctgaac aacaagatta
     7021 atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat gtaccgcgcg
     7081 gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg gtttcgtgtg
     7141 ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac cacgactcca
     7201 agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat aaaccaccaa
     7261 actgccaaaa aatgaaaact gtggacaagc tctgtccgtt tgctggcaac tgcaagggtc
     7321 tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat ttgtttttta
     7381 ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa ttgaaaacgc
     7441 gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac agttaatttg
     7501 cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt cttcgtattc
     7561 cttctctttt tcatttttct cctcataaaa attaacatag ttattatcgt atccatatat
     7621 gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt ttttaatggg
     7681 gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat tttctggtag
     7741 ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt tgggatcgtc
     7801 ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa ttacaccatt
     7861 ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt taaacaaaaa
     7921 cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt taaaaataac
     7981 agccattgta atgagacgca caaactaata tcaca
//
