LOCUS       sequence-436842-        8978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eIF4G-Sk protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8978)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8978)
  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-eIF4G-Sk
FEATURES             Location/Qualifiers
     source          1..8978
                     /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    4445..4579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4482..4501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4536..4555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4652..5024
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(5376..5393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5486..6074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5566..5585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6245..7036,7037..7105))
                     /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     6868..6887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7106..7210)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7511..7533
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7525..7542
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7526..7542
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7553..7572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7553..7571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8182..8306
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(8344..8949)
                     /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 actgatgaaa cagcacaacc tacccagagc gcacccaggc aagaatccgc
     1501 agccccgaaa cagacggacg acgaccaaca ggtttcacaa cagcaacaac agaggggtta
     1561 caacaattat aatgacggct caaattacaa ccaaaaaaag ccatataata gtaacagacc
     1621 ccatcagcag cgtggcggca agtttggccc aaacagatat aattcaagag gtaattataa
     1681 cggaggtgga aactataggg gcggacacat gggagctaac ggatcgaatg ttccatggac
     1741 gggttattac aataattatc ctgtctacta ccaacctcaa cagatgggtg cgggaaacgc
     1801 aatcccagtt aatgtagccc ccgcagatga aaaatcttct ccggtgccta ccaagattga
     1861 gattaccaca aagtctggag agcatctcga tcttaaagag caacataagg tgaaattgca
     1921 gtctaaagaa acctctactg caccaactcc cctggatccc aagtcggaag aagccagcga
     1981 ctcggcacct acgcctacgc ctacacctgc aaccacagac tcgaaagagt cttcagaaga
     2041 gaacatatca gaggcagaga agacgagaag gaattttatt gaacaggtaa agcttcgtaa
     2101 ggctgcgttg gaaaagaagc gtaaagaaca attggagggc gctggtggtt cgtccaacgt
     2161 atcaactgag accgctcctg tcaaagccga agaaacagag cccgtgatct ctgaagcggg
     2221 tgaaaaaaag ccggccgagc tggagacaaa ccaggacaca ccttcggaag aaaacggcca
     2281 ggaggagcag ggtcagataa aagaggagag tacaacgaag gcgctgacat tcgctgaaag
     2341 acttaagctc aagaaacaac agaaggaaca ggagaagact gaagagaaag aaaatcaaga
     2401 agagcctgtg caagaagagg cgaattccgt gactaaaaca gcgaccgtgc cgtccacaga
     2461 gcaagatcaa gaacagacag aaacaagtga gacggaacgc agcgaggtag aagaaagcac
     2521 accacaaacc gccgagccaa cagccacaat cccgggaacc ctcgcggaat ctggcgaggc
     2581 ggacgacacg gaagtcaaag tcgagtcaga ggccgaggta gaggcagaca tagagacagc
     2641 ggctgacgat gaagcagaca acggaacaaa tacagttagc cacgtgctga atgtcctgaa
     2701 ggacgccagg cccatagaag atgtgttttc atttatctat ccagaaggta ttgagggtcc
     2761 tgacattaaa tataaaaaag aacacgtgaa gtatacttac ggtccaacgt tcctccttca
     2821 gttcaaggac aaactgaacg taaaagcaga tgcagagtgg gtacagtcaa cagcgagtaa
     2881 gattgtcata cccccgggca tgggccgtgg aaataagtct agggatccgt caagatttag
     2941 caataactcg tccaggggtc acgacttcag gagcaactca gtacgcaata tggatgacag
     3001 agcaaacagt agaacctctt cgaagcgtcg ttccaaacgt atgaacgacg ataggcgctc
     3061 aaaccgcagc tacacaagca gaagagatag ggagaggggc tcctaccgca atgaggataa
     3121 gcgcgatgac gataggcccc gtgaagaagt ggccccactc gttccatcta gcaatcgttg
     3181 gatacctaaa tccaaagcca agtccaccga aaaaaagctt gcgcctgacg gaaagaccga
     3241 gcttttggac aaggatgaag tcgagagaaa gatgaaaagc cttttgaaca agctgacttt
     3301 ggagatgttt gatgctatca gttctgagat tttggctatt gcgaacatct ccgtctggga
     3361 aacgaacggc gagactctga aagcggtgat agaacaagta tttctgaaag cgtgcgacga
     3421 acctcattgg tcctcaatgt acgcacaatt gtgcggaaaa gtcgttaaag agcttaatcc
     3481 tgacataagc gatgaaacga acgaaggcaa gactggtccg aaacttgtat tgcattatct
     3541 ggttgccagg tgtcatgcgg aatttgacaa gggctggaca gacaagctgc cgaccaaaga
     3601 agacggcacg ccgctcgagc ctgaaatgat gtccgaagag tactatgcag ctgcatcggc
     3661 aaaaagaagg ggtcttggct tggttagatt tatcggattc ctttaccgtt tgaacctctt
     3721 gacgggtaaa atgatgttcg aatgctttag acgtctgatg aaagatttga cagattcccc
     3781 gtcagaggag acgctcgagt cggtcattga gctgctgaac accgtaggag aacaatttga
     3841 gacagactcg tttagaaccg gccaggctac tctcgaagga tctcagctct tggactcgct
     3901 gtttggcata ctggacaacg ttatacagac ggctgagatg agttctcgta ttaaattcaa
     3961 gttggtagac ataaaggagc tgcgtcacga caagaactgg aatagtgaca aaaaagacga
     4021 tggcccaaag accattcagc aaatccacga agaggaggaa cgccagaggc agcttaagaa
     4081 taattcaagg agtaattcac gccgcaccaa caattcatcc aataggcaca gtttcagaag
     4141 ggatgcgcct cccgctagca aagacagctt tatagcgaca agaacatata gtcagcgcaa
     4201 tacacagcgt gcgcctccac cgcgcgagga gccttccgct cctacctcca cagcaacaaa
     4261 tatgttcagc gctctcatgg gagaatcgga cgacgaggaa ggcgcgccgt aatagaagct
     4321 tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4381 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4441 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4561 atcttatcat gtctggatct gatcactgct tgagcctagg agatccgaac cagataagtg
     4621 aaagcctgca tccgcagatc ttaattaata aaacacgata cattgttatt agtacattta
     4681 ttaagcgcta gattctgtgc gttgttgatt tacagacaat tgttgtacgt attttaataa
     4741 ttcattaaat ttataatctt tagggtggta tgttagagcg aaaatcaaat gattttcagc
     4801 gtctttatat ctgaatttaa atattaaatc ctcaatagat ttgtaaaata ggtttcgatt
     4861 agtttcaaac aagggttgtt tttccgaacc gatggctgga ctatctaatg gattttcgct
     4921 caacgccaca aaacttgcca aatcttgtag cagcaatcta gctttgtcga tattcgtttg
     4981 tgttttgttt tgtaataaag gttcgacgtc gttcaaaata ttatgcgctt ttgtatttct
     5041 ttcatcactg tcgttagtgt acaattgact cgacgtaaac acgttaaata acgcttggac
     5101 atatttaaca tcgggcgtgt tagctttatt aggccgatta tcgtcgtcgt cccaaccctc
     5161 gtcgttagaa gttgcttccg aagacgattt tgccatagcc acacgacgcc tattaattgt
     5221 gtcggctaac acgtccgcga tcaaatttgt agttgagctt tttgggcctc atgggccttc
     5281 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat
     5341 agctgtttcc ttgcgtattg ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg
     5401 gtcgttcggg taaagcctgg ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc
     5461 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5521 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5581 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5641 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5701 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5761 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5821 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5881 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5941 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6001 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6061 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6241 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6301 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6361 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa
     6421 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6481 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6541 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6601 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6661 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6721 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6781 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6841 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6901 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6961 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7021 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7081 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7141 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7201 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     7261 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7321 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtggcc gctacagggc
     7381 gctcccattc gccattcagg ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct
     7441 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7501 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcga cgtaatacga
     7561 ctcactatag ggcgaattgg cggaaggccg tcaaggccgt gacgttaaaa ctattaagcc
     7621 atccaatcga ccgttagtcg aatcaggacc gctggtgcga gaagccgcga agtatggcga
     7681 atgcatcgta taacgtgtgg agtccgctca ttagagcgtc atgtttagac aagaaagcta
     7741 catatttaat tgatcccgat gattttattg ataaattgac cctaactcca tacacggtat
     7801 tctacaatgg cggggttttg gtcaaaattt ccggactgcg attgtacatg ctgttaacgg
     7861 ctccgcccac tattaatgaa attaaaaatt ccaattttaa aaaacgcagc aagagaaaca
     7921 tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt ggacatgctg aacaacaaga
     7981 ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt gaaagaaaac aatgtaccgc
     8041 gcggcggtat gtacaggaag aggtttatac taaactgtta cattgcaaac gtggtttcgt
     8101 gtgccaagtg tgaaaaccga tgtttaatca aggctctgac gcatttctac aaccacgact
     8161 ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc ccaagatgtg tataaaccac
     8221 caaactgcca aaaaatgaaa actgtggaca agctctgtcc gtttgctggc aactgcaagg
     8281 gtctcaatcc tatttgtaat tattgaataa taaaacaatt ataaatgcta aatttgtttt
     8341 ttattaacga tacaaaccaa acgcaacaag aacatttgta gtattatcta taattgaaaa
     8401 cgcgtagtta taatcgctga ggtaatattt aaaatcattt tcaaatgatt cacagttaat
     8461 ttgcgacaat ataattttat tttcacataa actagacgcc ttgtcgtctt cttcttcgta
     8521 ttccttctct ttttcatttt tctcctcata aaaattaaca tagttattat cgtatccata
     8581 tatgtatcta tcgtatagag taaatttttt gttgtcataa atatatatgt cttttttaat
     8641 ggggtgtata gtaccgctgc gcatagtttt tctgtaattt acaacagtgc tattttctgg
     8701 tagttcttcg gagtgtgttg ctttaattat taaatttata taatcaatga atttgggatc
     8761 gtcggttttg tacaatatgt tgccggcata gtacgcagct tcttctagtt caattacacc
     8821 attttttagc agcaccggat taacataact ttccaaaatg ttgtacgaac cgttaaacaa
     8881 aaacagttca cctccctttt ctatactatt gtctgcgagc agttgtttgt tgttaaaaat
     8941 aacagccatt gtaatgagac gcacaaacta atatcaca
//
