LOCUS       sequence-436799-        8981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eIF4G-213 protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8981)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8981)
  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-213
FEATURES             Location/Qualifiers
     source          1..8981
                     /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    4448..4582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4485..4504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4539..4558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4655..5027
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(5379..5396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5489..6077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5569..5588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6248..7039,7040..7108))
                     /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     6871..6890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7109..7213)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7514..7536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7528..7545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7529..7545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7556..7575
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7556..7574
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8185..8309
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(8347..8952)
                     /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 actgacgaaa cagctcaccc gacacagagc gcctcgaaac aggagagtgc
     1501 agctttgaag cagaccggcg atgaccagca ggagtcccag cagcagagag gttatacaaa
     1561 ctataacaac ggaagcaact acacccagaa aaagccatat aacagtaaca gaccacacca
     1621 gcagagagga ggtaaattcg gacccaaccg ctataacaac agaggaaatt ataacggagg
     1681 aggctcattt agaggaggcc atatgggtgc gaactcatca aatgtgcctt ggaccggata
     1741 ttacaataat tatcctgtat actatcagcc gcaacagatg gccgcagcag gaagcgctcc
     1801 tgcgaaccct attcctgtag aagaaaagag tcccgtcccc acgaaaattg aaataacaac
     1861 caagtcaggc gagcaccttg atctgaaaga acagcacaag gcaaaacttc aatcccaaga
     1921 gaggagcact gtttctcccc aaccagaatc caaacttaaa gaaacgagcg atagtacatc
     1981 taccagtacg ccgactccca ctccgtccac gaatgatagc aaagctagca gcgaagaaaa
     2041 tatctccgag gcagaaaaaa ctcgtaggaa tttcatcgag caggcagcag cagcaaaggc
     2101 ggcgcttgaa aaaaagagga aagagcaatt ggagggaagt tcgggtaaca acaacatccc
     2161 gatgaagacc actccagaaa acgtagaaga aaaaggatca gataaacccg aggtcactga
     2221 aaaaaccaag cctgcagagg agaagtccgc ggaacctgag gtcaaacaag aaactcccgc
     2281 agaggaaggc gaacaaggcg agaaaggtca gattaaagaa gagagtacgc ctaaagttct
     2341 gactttcgcg gaacgtctta agctgaagaa gcagcagaag gaaagagagg aaaagacaga
     2401 gggtaaggag aataaggaag tgcctgttca agaagagaca aagtccgcca tagaaagtgc
     2461 gcctgtgcca ccttcagaac aagtaaaaga agagacggag gttgctgaga cggaacaatc
     2521 caatatcgat gagtcagcca ctaccccggc tattccgaca aaaagcgatg aagctgaggc
     2581 agaggtggaa gctgaagcag gagacgcggg tacaaaaatc ggactggaag ccgagattga
     2641 gaccacaacc gacgagactg acgatggcac caacacagtc agtcatattt tgaatgttct
     2701 taaagacgct acgccaattg aagatgtctt ctcattcaat tacccagaag gtatcgaagg
     2761 tccagacata aaatataaaa aggagcatgt aaagtataca tatggcccaa ctttcttgtt
     2821 gcaattcaag gacaaactta acgtcaaggc tgacgcagaa tgggtccaaa gtactgctag
     2881 taaaatcgta attcccccag gcatgggccg tggtaatcgc agccgcgaca gtggacgttt
     2941 tggaaacaac tcaagccgcg gccacgactt taggaacact tcggtacgca acatggatga
     3001 ccgcgctaat tcccgtacga gcagcaagcg taggtcgaag cgcatgaatg acgaccgcag
     3061 gagcaataga tcatacacca gccgcagaga tagggaaagg ggttcttatc gcaacgaaga
     3121 gaagcgtgaa gacgacaagc caaaggaaga agtcgctccc cttgtaccgt cagctaacag
     3181 gtgggtaccg aagttcaaat cgaagaaaac tgagaaaaaa cttgcccctg atggcaaaac
     3241 ggaactgctc gataaagatg aagtggagcg caagatgaag tcacttttga acaagttgac
     3301 tctggagatg tttgatgcca ttagttctga aatcctggca atcgccaaca tcagcgtatg
     3361 ggagacgaac ggcgagacgc ttaaggcggt aatagaacaa atctttctca aagcgtgtga
     3421 cgagccgcat tggagctcca tgtatgctca gctgtgcggc aaagtagtga aagaactcaa
     3481 tccagacata acagatgaaa ccaacgaggg aaagactggt ccgaagttgg tcttgcacta
     3541 cctggtagct cgctgtcatg ctgaattcga taaaggttgg actgataagc ttccgacgaa
     3601 tgaagacgga accccgcttg aacccgagat gatgtctgag gagtactatg ctgccgcgtc
     3661 cgccaagagg cgcggtctgg gacttgtccg ctttattggc ttcctctatc gtcttaacct
     3721 tcttacaggc aaaatgatgt ttgaatgttt taggagactg atgaaggacc tcactgatag
     3781 tcctagtgag gagacgctgg aatccgttgt tgagctcctg aataccgtag gagagcagtt
     3841 cgagactgac tctttcagga ccggacaggc gactcttgaa ggctcgcagt tgctcgactc
     3901 gttgtttgga atcttggata atattataca aacagccaag ataagcagta gaataaaatt
     3961 taaacttata gatattaagg agctcaggca cgacaaaaat tggaatagtg acaaaaaaga
     4021 taacggaccg aaaactatac aacagattca cgaggaagaa gagaggcaaa ggcagcttaa
     4081 gaacaactct aggagcaact caagacgcac aaacaattcg tccaaccgtc acagcttccg
     4141 cagagacgct ccgccagcct cgaaggattc gtttataacc acaagaactt atagccagcg
     4201 caacagccaa agagcgccac cgccaaagga agaaccagct gccccaacaa gcacagctac
     4261 gaacatgttc tctgcgctca tgggtgagtc cgatgatgaa gaaggcgcgc cgtaatagaa
     4321 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
     4381 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4441 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4501 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4561 tgtatcttat catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa
     4621 gtgaaagcct gcatccgcag atcttaatta ataaaacacg atacattgtt attagtacat
     4681 ttattaagcg ctagattctg tgcgttgttg atttacagac aattgttgta cgtattttaa
     4741 taattcatta aatttataat ctttagggtg gtatgttaga gcgaaaatca aatgattttc
     4801 agcgtcttta tatctgaatt taaatattaa atcctcaata gatttgtaaa ataggtttcg
     4861 attagtttca aacaagggtt gtttttccga accgatggct ggactatcta atggattttc
     4921 gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat ctagctttgt cgatattcgt
     4981 ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa atattatgcg cttttgtatt
     5041 tctttcatca ctgtcgttag tgtacaattg actcgacgta aacacgttaa ataacgcttg
     5101 gacatattta acatcgggcg tgttagcttt attaggccga ttatcgtcgt cgtcccaacc
     5161 ctcgtcgtta gaagttgctt ccgaagacga ttttgccata gccacacgac gcctattaat
     5221 tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag ctttttgggc ctcatgggcc
     5281 ttccgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaacatggt
     5341 catagctgtt tccttgcgta ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc
     5401 tcggtcgttc gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa aaggccagga
     5461 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5521 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5581 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5641 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5701 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5761 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5821 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5881 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5941 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6001 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6061 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6121 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6181 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6241 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6301 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6361 ctggccccag tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag
     6421 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6481 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6541 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6601 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6661 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6721 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6781 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6841 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6901 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6961 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7021 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7081 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7141 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7201 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     7261 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     7321 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg gccgctacag
     7381 ggcgctccca ttcgccattc aggctgcgca actgttggga agggcgtttc ggtgcgggcc
     7441 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     7501 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgacgtaata
     7561 cgactcacta tagggcgaat tggcggaagg ccgtcaaggc cgtgacgtta aaactattaa
     7621 gccatccaat cgaccgttag tcgaatcagg accgctggtg cgagaagccg cgaagtatgg
     7681 cgaatgcatc gtataacgtg tggagtccgc tcattagagc gtcatgttta gacaagaaag
     7741 ctacatattt aattgatccc gatgatttta ttgataaatt gaccctaact ccatacacgg
     7801 tattctacaa tggcggggtt ttggtcaaaa tttccggact gcgattgtac atgctgttaa
     7861 cggctccgcc cactattaat gaaattaaaa attccaattt taaaaaacgc agcaagagaa
     7921 acatttgtat gaaagaatgc gtagaaggaa agaaaaatgt cgtggacatg ctgaacaaca
     7981 agattaatat gcctccgtgt ataaaaaaaa tattgaacga tttgaaagaa aacaatgtac
     8041 cgcgcggcgg tatgtacagg aagaggttta tactaaactg ttacattgca aacgtggttt
     8101 cgtgtgccaa gtgtgaaaac cgatgtttaa tcaaggctct gacgcatttc tacaaccacg
     8161 actccaagtg tgtgggtgaa gtcatgcatc ttttaatcaa atcccaagat gtgtataaac
     8221 caccaaactg ccaaaaaatg aaaactgtgg acaagctctg tccgtttgct ggcaactgca
     8281 agggtctcaa tcctatttgt aattattgaa taataaaaca attataaatg ctaaatttgt
     8341 tttttattaa cgatacaaac caaacgcaac aagaacattt gtagtattat ctataattga
     8401 aaacgcgtag ttataatcgc tgaggtaata tttaaaatca ttttcaaatg attcacagtt
     8461 aatttgcgac aatataattt tattttcaca taaactagac gccttgtcgt cttcttcttc
     8521 gtattccttc tctttttcat ttttctcctc ataaaaatta acatagttat tatcgtatcc
     8581 atatatgtat ctatcgtata gagtaaattt tttgttgtca taaatatata tgtctttttt
     8641 aatggggtgt atagtaccgc tgcgcatagt ttttctgtaa tttacaacag tgctattttc
     8701 tggtagttct tcggagtgtg ttgctttaat tattaaattt atataatcaa tgaatttggg
     8761 atcgtcggtt ttgtacaata tgttgccggc atagtacgca gcttcttcta gttcaattac
     8821 accatttttt agcagcaccg gattaacata actttccaaa atgttgtacg aaccgttaaa
     8881 caaaaacagt tcacctccct tttctatact attgtctgcg agcagttgtt tgttgttaaa
     8941 aataacagcc attgtaatga gacgcacaaa ctaatatcac a
//
