LOCUS       sequence-436882-        8069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eIF4A-mCherry protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8069)
  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: pOCC240-MBP-eIF4A-mCherry
FEATURES             Location/Qualifiers
     source          1..8069
                     /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             2615..2635
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2636..3343
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2781..2799)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3049..3068
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3344..3367
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3377..3394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3536..3670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3573..3592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3627..3646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3743..4115
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4467..4484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4577..5165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4657..4676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5336..6127,6128..6196))
                     /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     5959..5978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6197..6301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6602..6624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6616..6633
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6617..6633
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6644..6663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6644..6662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7273..7397
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7435..8040)
                     /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 aatgtcggaa ggtataaccg atattgagga
     1441 gagtcagata caaacaaatt atgacaaggt ggtttataaa ttcgacgata tggaattgga
     1501 cgagaacttg ttgcgtggcg tgtttggata tggctttgag gagccgtccg cgattcaaca
     1561 gagggccata atgccgatta tcgaaggaca cgacgtactg gctcaagccc aaagcggaac
     1621 gggtaaaaca ggaacttttt ctattgccgc tttgcagcgt atcgatacct cagtcaaagc
     1681 cccccaagcg ttgatgttgg cgcctactag agagttggcg ttgcagatac aaaaagttgt
     1741 catggcactc gcatttcaca tggatatcaa agtacacgcc tgcattggag gaacgtcatt
     1801 tgtagaggac gcggaaggac tgagagacgc acagattgtt gtcggtacac ccggtcgtgt
     1861 attcgataat attcaacgtc gcagattccg cacggataag ataaagatgt ttattcttga
     1921 tgaggctgat gaaatgctct catccggatt taaggaacaa atatatcaaa ttttcacctt
     1981 gttgccacca acaacgcagg tggttctgct ctccgcgact atgccgaatg acgtcctgga
     2041 ggttaccaca aagttcatgc gtaatcctgt ccgcatcctc gtcaagaaag acgagcttac
     2101 tctggaagga attaaacagt tctatgtaaa tgtagaggag gaagaataca agtacgaatg
     2161 tctgactgat ttgtacgatt caatatccgt cactcaagcc gttatctttt gcaacactag
     2221 aagaaaagtt gaggaactga caacaaagct tcgcaacgat aagttcacag tctctgccat
     2281 atattcagac ttgccccaac aagaacgcga caccataatg aaagaattta gaagcggttc
     2341 gtccaggatt ctcatatcta ccgacttgtt ggccaggggc attgatgtgc agcaggtaag
     2401 tctcgtgatc aactatgatc tgccggccaa caaagaaaac tatatacatc gtataggacg
     2461 tggaggccgt tttggacgca agggcgtagc aattaatttc gtcactaacg aagacgtagg
     2521 tgccatgagg gagcttgaga aattttattc cacccagatc gaagagctgc cgtcagatat
     2581 tgctacactg ttgaatggcg cgccgggatc cagcgaaaac ttgtatttcc agggcatggt
     2641 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     2701 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2761 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     2821 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     2881 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2941 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     3001 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     3061 gaagaagacg atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     3121 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     3181 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     3241 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     3301 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagctggaag ttctgttcca
     3361 ggggccgggc agcagccatc accatcacca tcacagtgga taatagaagc ttgtcgagaa
     3421 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3481 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3541 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3601 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3661 tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaagcctgc
     3721 atccgcagat cttaattaat aaaacacgat acattgttat tagtacattt attaagcgct
     3781 agattctgtg cgttgttgat ttacagacaa ttgttgtacg tattttaata attcattaaa
     3841 tttataatct ttagggtggt atgttagagc gaaaatcaaa tgattttcag cgtctttata
     3901 tctgaattta aatattaaat cctcaataga tttgtaaaat aggtttcgat tagtttcaaa
     3961 caagggttgt ttttccgaac cgatggctgg actatctaat ggattttcgc tcaacgccac
     4021 aaaacttgcc aaatcttgta gcagcaatct agctttgtcg atattcgttt gtgttttgtt
     4081 ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct tttgtatttc tttcatcact
     4141 gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat aacgcttgga catatttaac
     4201 atcgggcgtg ttagctttat taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga
     4261 agttgcttcc gaagacgatt ttgccatagc cacacgacgc ctattaattg tgtcggctaa
     4321 cacgtccgcg atcaaatttg tagttgagct ttttgggcct catgggcctt ccgctcactg
     4381 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca tagctgtttc
     4441 cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4501 gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4561 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4621 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4681 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4741 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4801 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4861 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4921 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4981 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5041 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5101 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5161 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5221 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5281 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5341 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5401 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5461 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     5521 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5581 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5641 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5701 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5761 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5821 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5881 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5941 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6001 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6061 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6121 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6181 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6241 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6301 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     6361 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     6421 ttataaatca aaagaataga ccgagatagg gttgagtggc cgctacaggg cgctcccatt
     6481 cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc ttcgctatta
     6541 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6601 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg actcactata
     6661 gggcgaattg gcggaaggcc gtcaaggccg tgacgttaaa actattaagc catccaatcg
     6721 accgttagtc gaatcaggac cgctggtgcg agaagccgcg aagtatggcg aatgcatcgt
     6781 ataacgtgtg gagtccgctc attagagcgt catgtttaga caagaaagct acatatttaa
     6841 ttgatcccga tgattttatt gataaattga ccctaactcc atacacggta ttctacaatg
     6901 gcggggtttt ggtcaaaatt tccggactgc gattgtacat gctgttaacg gctccgccca
     6961 ctattaatga aattaaaaat tccaatttta aaaaacgcag caagagaaac atttgtatga
     7021 aagaatgcgt agaaggaaag aaaaatgtcg tggacatgct gaacaacaag attaatatgc
     7081 ctccgtgtat aaaaaaaata ttgaacgatt tgaaagaaaa caatgtaccg cgcggcggta
     7141 tgtacaggaa gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt
     7201 gtgaaaaccg atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg
     7261 tgggtgaagt catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc
     7321 aaaaaatgaa aactgtggac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc
     7381 ctatttgtaa ttattgaata ataaaacaat tataaatgct aaatttgttt tttattaacg
     7441 atacaaacca aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt
     7501 ataatcgctg aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa
     7561 tataatttta ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc
     7621 tttttcattt ttctcctcat aaaaattaac atagttatta tcgtatccat atatgtatct
     7681 atcgtataga gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat
     7741 agtaccgctg cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc
     7801 ggagtgtgtt gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt
     7861 gtacaatatg ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag
     7921 cagcaccgga ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc
     7981 acctcccttt tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat
     8041 tgtaatgaga cgcacaaact aatatcaca
//
