LOCUS       sequence-436836-        8705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ded1p-monoGFP protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8705)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8705)
  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"
COMMENT     Sequence Label: pOCC120-MBP-Ded1-monoGFP
FEATURES             Location/Qualifiers
     source          1..8705
                     /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             224..1324
                     /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     1298..1321
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1379..1402
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3263..3265,3266..3268,3269..3979)
                     /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(3308..3329)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3569..3588)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3916..3937
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3980..4003
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4013..4030
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4172..4306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4209..4228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4263..4282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4379..4751
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(5103..5120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5213..5801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5293..5312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5972..6763,6764..6832))
                     /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     6595..6614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6833..6937)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7238..7260
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7252..7269
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7253..7269
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7280..7299
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7280..7298
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7909..8033
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(8071..8676)
                     /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 ggcatgaaaa tcgaagaagg
      241 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      301 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      361 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      421 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      481 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      541 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      601 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      661 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      721 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      781 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      841 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      901 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      961 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
     1021 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1081 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1141 gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca
     1201 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1261 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1321 gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca gcggcagact
     1381 ggaagttctg ttccaggggc cggcggccgc aatggctgaa ctgtcagaac aggtacaaaa
     1441 cctgagtatc aatgataaca atgaaaatgg atacgttccg ccgcatttgc gcggaaagcc
     1501 gagatctgca cgcaataact catccaatta taacaacaac aatggtggtt acaatggtgg
     1561 aagaggcgga ggttcctttt tttcaaacaa tcgccgtggc ggatatggta acggtggatt
     1621 ttttggcggt aataatggcg gatccaggag taatggcaga tcaggtggtc gttggatcga
     1681 tggcaagcac gttcctgcgc cacgtaacga gaaggctgaa attgcgatct tcggagttcc
     1741 cgaggatcca aatttccaat catccggaat taactttgac aactacgatg acatcccggt
     1801 tgatgcctca ggaaaagatg tccctgaacc gataaccgaa tttacgtctc cccctttgga
     1861 cggtttgttg ctggagaaca tcaaactcgc tcgttttacc aagccaacac ccgtacagaa
     1921 atattctgtc ccgattgtag ctaacggtag agacttgatg gcatgcgctc aaacaggatc
     1981 cggcaagacg ggcggatttc tgtttcctgt gctgtctgaa tcattcaaaa ctggacccag
     2041 cccccaaccc gaaagtcagg gctcatttta ccaaagaaag gcgtacccga cagctgtgat
     2101 tatggctccc acgcgcgagc tcgcaactca gatattcgac gaggcgaaaa aatttacata
     2161 cagatcatgg gttaaggcat gcgtagtata cggcggtagt cctattggaa accagctcag
     2221 agagatagag agaggctgtg accttctggt ggcgactccc ggacgcctga atgatttgct
     2281 tgaaagggga aaaataagtc tcgcaaacgt gaaatacctt gtacttgatg aggctgacag
     2341 gatgctcgat atgggctttg aacctcaaat caggcatatt gtcgaagatt gcgacatgac
     2401 gcccgttgga gagagacaaa cattgatgtt cagtgcaaca tttccagcag acatccagca
     2461 tcttgcccgt gatttcctgt ccgattacat ctttctgtca gtcggcagag ttggctccac
     2521 gtctgagaac atcacacaga aggttttgta cgtcgagaat caagataaaa aatcagcttt
     2581 gttggacttg ttgagtgcat cgaccgacgg tctgacactt atttttgtag aaactaagag
     2641 aatggctgat caacttacag acttcttgat tatgcagaac tttcgcgcca cagccatcca
     2701 cggtgatcgc acacaatccg agcgcgagag ggcattggcg gcctttcgca gcggcgcggc
     2761 tactctgttg gtagctacag ctgtcgctgc tcgcggattg gacattccta atgtcaccca
     2821 tgttataaat tatgatttgc catccgatgt agacgattat gtgcatcgta tcggcagaac
     2881 tggaagagcc ggaaatacgg gactcgccac tgcatttttt aattcagaaa actccaatat
     2941 tgtgaagggc ctccacgaga tcctgacaga ggctaaccaa gaagtgccgt ctttcctgaa
     3001 ggatgctatg atgagcgccc ccggatcgcg ttctaacagt aggcgcggag gtttcggacg
     3061 caataacaat agagactacc gtaaagcagg aggtgcgtcc gcaggaggat ggggatcaag
     3121 tcgttctagg gataactcct ttaggggcgg tagtggttgg ggtagtgatt ccaagagttc
     3181 aggttggggt aattcgggcg gatccaacaa ctcttcatgg tggggcgcgc cgggatccgc
     3241 tggctccgct gctggttctg gcatggtgag caagggcgag gagctgttca ccggggtggt
     3301 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3361 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3421 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3481 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3541 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3601 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3661 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3721 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3781 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3841 cgtgctgctg cccgacaacc actacctgag cacccagtcc aaactgagca aagaccccaa
     3901 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3961 catggacgag ctgtacaagc tggaagttct gttccagggg ccgggcagca gccatcacca
     4021 tcaccatcac agtggataat agaagcttgt cgagaagtac tagaggatca taatcagcca
     4081 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4141 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4201 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4261 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga
     4321 gcctaggaga tccgaaccag ataagtgaaa gcctgcatcc gcagatctta attaataaaa
     4381 cacgatacat tgttattagt acatttatta agcgctagat tctgtgcgtt gttgatttac
     4441 agacaattgt tgtacgtatt ttaataattc attaaattta taatctttag ggtggtatgt
     4501 tagagcgaaa atcaaatgat tttcagcgtc tttatatctg aatttaaata ttaaatcctc
     4561 aatagatttg taaaataggt ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat
     4621 ggctggacta tctaatggat tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag
     4681 caatctagct ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt
     4741 caaaatatta tgcgcttttg tatttctttc atcactgtcg ttagtgtaca attgactcga
     4801 cgtaaacacg ttaaataacg cttggacata tttaacatcg ggcgtgttag ctttattagg
     4861 ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag acgattttgc
     4921 catagccaca cgacgcctat taattgtgtc ggctaacacg tccgcgatca aatttgtagt
     4981 tgagcttttt gggcctcatg ggccttccgc tcactgcccg ctttccagtc gggaaacctg
     5041 tcgtgccagc tgcattaaca tggtcatagc tgtttccttg cgtattgggc gctctccgct
     5101 tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa agcctggggt gcctaatgag
     5161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac
     6121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6961 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7021 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7081 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
     7141 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7201 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     7261 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
     7321 aggccgtgac gttaaaacta ttaagccatc caatcgaccg ttagtcgaat caggaccgct
     7381 ggtgcgagaa gccgcgaagt atggcgaatg catcgtataa cgtgtggagt ccgctcatta
     7441 gagcgtcatg tttagacaag aaagctacat atttaattga tcccgatgat tttattgata
     7501 aattgaccct aactccatac acggtattct acaatggcgg ggttttggtc aaaatttccg
     7561 gactgcgatt gtacatgctg ttaacggctc cgcccactat taatgaaatt aaaaattcca
     7621 attttaaaaa acgcagcaag agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa
     7681 atgtcgtgga catgctgaac aacaagatta atatgcctcc gtgtataaaa aaaatattga
     7741 acgatttgaa agaaaacaat gtaccgcgcg gcggtatgta caggaagagg tttatactaa
     7801 actgttacat tgcaaacgtg gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg
     7861 ctctgacgca tttctacaac cacgactcca agtgtgtggg tgaagtcatg catcttttaa
     7921 tcaaatccca agatgtgtat aaaccaccaa actgccaaaa aatgaaaact gtggacaagc
     7981 tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat ttgtaattat tgaataataa
     8041 aacaattata aatgctaaat ttgtttttta ttaacgatac aaaccaaacg caacaagaac
     8101 atttgtagta ttatctataa ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa
     8161 atcattttca aatgattcac agttaatttg cgacaatata attttatttt cacataaact
     8221 agacgccttg tcgtcttctt cttcgtattc cttctctttt tcatttttct cctcataaaa
     8281 attaacatag ttattatcgt atccatatat gtatctatcg tatagagtaa attttttgtt
     8341 gtcataaata tatatgtctt ttttaatggg gtgtatagta ccgctgcgca tagtttttct
     8401 gtaatttaca acagtgctat tttctggtag ttcttcggag tgtgttgctt taattattaa
     8461 atttatataa tcaatgaatt tgggatcgtc ggttttgtac aatatgttgc cggcatagta
     8521 cgcagcttct tctagttcaa ttacaccatt ttttagcagc accggattaa cataactttc
     8581 caaaatgttg tacgaaccgt taaacaaaaa cagttcacct cccttttcta tactattgtc
     8641 tgcgagcagt tgtttgttgt taaaaataac agccattgta atgagacgca caaactaata
     8701 tcaca
//
