LOCUS       sequence-436878-        7310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eIF4A protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7310)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 7310)
  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: pOCC102-MBP-eIF4A
FEATURES             Location/Qualifiers
     source          1..7310
                     /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"
     polyA_signal    2777..2911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2814..2833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2868..2887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2984..3356
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3708..3725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3818..4406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3898..3917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4577..5368,5369..5437))
                     /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     5200..5219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5438..5542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5843..5865
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5857..5874
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5858..5874
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5885..5904
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5885..5903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6514..6638
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6676..7281)
                     /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 tcggaaggta taaccgatat tgaggagagt cagatacaaa caaattatga
     1501 caaggtggtt tataaattcg acgatatgga attggacgag aacttgttgc gtggcgtgtt
     1561 tggatatggc tttgaggagc cgtccgcgat tcaacagagg gccataatgc cgattatcga
     1621 aggacacgac gtactggctc aagcccaaag cggaacgggt aaaacaggaa ctttttctat
     1681 tgccgctttg cagcgtatcg atacctcagt caaagccccc caagcgttga tgttggcgcc
     1741 tactagagag ttggcgttgc agatacaaaa agttgtcatg gcactcgcat ttcacatgga
     1801 tatcaaagta cacgcctgca ttggaggaac gtcatttgta gaggacgcgg aaggactgag
     1861 agacgcacag attgttgtcg gtacacccgg tcgtgtattc gataatattc aacgtcgcag
     1921 attccgcacg gataagataa agatgtttat tcttgatgag gctgatgaaa tgctctcatc
     1981 cggatttaag gaacaaatat atcaaatttt caccttgttg ccaccaacaa cgcaggtggt
     2041 tctgctctcc gcgactatgc cgaatgacgt cctggaggtt accacaaagt tcatgcgtaa
     2101 tcctgtccgc atcctcgtca agaaagacga gcttactctg gaaggaatta aacagttcta
     2161 tgtaaatgta gaggaggaag aatacaagta cgaatgtctg actgatttgt acgattcaat
     2221 atccgtcact caagccgtta tcttttgcaa cactagaaga aaagttgagg aactgacaac
     2281 aaagcttcgc aacgataagt tcacagtctc tgccatatat tcagacttgc cccaacaaga
     2341 acgcgacacc ataatgaaag aatttagaag cggttcgtcc aggattctca tatctaccga
     2401 cttgttggcc aggggcattg atgtgcagca ggtaagtctc gtgatcaact atgatctgcc
     2461 ggccaacaaa gaaaactata tacatcgtat aggacgtgga ggccgttttg gacgcaaggg
     2521 cgtagcaatt aatttcgtca ctaacgaaga cgtaggtgcc atgagggagc ttgagaaatt
     2581 ttattccacc cagatcgaag agctgccgtc agatattgct acactgttga atggcgcgcc
     2641 gtaatagaag cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga
     2701 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2761 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2821 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2881 actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta ggagatccga
     2941 accagataag tgaaagcctg catccgcaga tcttaattaa taaaacacga tacattgtta
     3001 ttagtacatt tattaagcgc tagattctgt gcgttgttga tttacagaca attgttgtac
     3061 gtattttaat aattcattaa atttataatc tttagggtgg tatgttagag cgaaaatcaa
     3121 atgattttca gcgtctttat atctgaattt aaatattaaa tcctcaatag atttgtaaaa
     3181 taggtttcga ttagtttcaa acaagggttg tttttccgaa ccgatggctg gactatctaa
     3241 tggattttcg ctcaacgcca caaaacttgc caaatcttgt agcagcaatc tagctttgtc
     3301 gatattcgtt tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa tattatgcgc
     3361 ttttgtattt ctttcatcac tgtcgttagt gtacaattga ctcgacgtaa acacgttaaa
     3421 taacgcttgg acatatttaa catcgggcgt gttagcttta ttaggccgat tatcgtcgtc
     3481 gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat tttgccatag ccacacgacg
     3541 cctattaatt gtgtcggcta acacgtccgc gatcaaattt gtagttgagc tttttgggcc
     3601 tcatgggcct tccgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3661 taacatggtc atagctgttt ccttgcgtat tgggcgctct ccgcttcctc gctcactgac
     3721 tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa
     3781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4501 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4561 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4621 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4681 gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca ccggctccag
     4741 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4801 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4861 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4921 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4981 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5041 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5101 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5161 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5221 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5281 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5341 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5401 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     5581 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5641 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgg
     5701 ccgctacagg gcgctcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgtttcg
     5761 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     5821 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     5881 gacgtaatac gactcactat agggcgaatt ggcggaaggc cgtcaaggcc gtgacgttaa
     5941 aactattaag ccatccaatc gaccgttagt cgaatcagga ccgctggtgc gagaagccgc
     6001 gaagtatggc gaatgcatcg tataacgtgt ggagtccgct cattagagcg tcatgtttag
     6061 acaagaaagc tacatattta attgatcccg atgattttat tgataaattg accctaactc
     6121 catacacggt attctacaat ggcggggttt tggtcaaaat ttccggactg cgattgtaca
     6181 tgctgttaac ggctccgccc actattaatg aaattaaaaa ttccaatttt aaaaaacgca
     6241 gcaagagaaa catttgtatg aaagaatgcg tagaaggaaa gaaaaatgtc gtggacatgc
     6301 tgaacaacaa gattaatatg cctccgtgta taaaaaaaat attgaacgat ttgaaagaaa
     6361 acaatgtacc gcgcggcggt atgtacagga agaggtttat actaaactgt tacattgcaa
     6421 acgtggtttc gtgtgccaag tgtgaaaacc gatgtttaat caaggctctg acgcatttct
     6481 acaaccacga ctccaagtgt gtgggtgaag tcatgcatct tttaatcaaa tcccaagatg
     6541 tgtataaacc accaaactgc caaaaaatga aaactgtgga caagctctgt ccgtttgctg
     6601 gcaactgcaa gggtctcaat cctatttgta attattgaat aataaaacaa ttataaatgc
     6661 taaatttgtt ttttattaac gatacaaacc aaacgcaaca agaacatttg tagtattatc
     6721 tataattgaa aacgcgtagt tataatcgct gaggtaatat ttaaaatcat tttcaaatga
     6781 ttcacagtta atttgcgaca atataatttt attttcacat aaactagacg ccttgtcgtc
     6841 ttcttcttcg tattccttct ctttttcatt tttctcctca taaaaattaa catagttatt
     6901 atcgtatcca tatatgtatc tatcgtatag agtaaatttt ttgttgtcat aaatatatat
     6961 gtctttttta atggggtgta tagtaccgct gcgcatagtt tttctgtaat ttacaacagt
     7021 gctattttct ggtagttctt cggagtgtgt tgctttaatt attaaattta tataatcaat
     7081 gaatttggga tcgtcggttt tgtacaatat gttgccggca tagtacgcag cttcttctag
     7141 ttcaattaca ccatttttta gcagcaccgg attaacataa ctttccaaaa tgttgtacga
     7201 accgttaaac aaaaacagtt cacctccctt ttctatacta ttgtctgcga gcagttgttt
     7261 gttgttaaaa ataacagcca ttgtaatgag acgcacaaac taatatcaca
//
