LOCUS       sequence-436750-        7517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eIF4G-RAR protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7517)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 7517)
  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-RAR
FEATURES             Location/Qualifiers
     source          1..7517
                     /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    2984..3118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3021..3040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3075..3094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3191..3563
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3915..3932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4025..4613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4105..4124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4784..5575,5576..5644))
                     /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     5407..5426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5645..5749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6050..6072
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6064..6081
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6065..6081
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6092..6111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6092..6110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6721..6845
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6883..7488)
                     /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 aatcgcagcc gcgacagtgg acgttttgga aacaactcaa gccgcggcca
     1501 cgactttagg aacacttcgg tacgcaacat ggatgaccgc gctaattccc gtacgagcag
     1561 caagcgtagg tcgaagcgca tgaatgacga ccgcaggagc aatagatcat acaccagccg
     1621 cagagatagg gaaaggggtt cttatcgcaa cgaagagaag cgtgaagacg acaagccaaa
     1681 ggaagaagtc gctccccttg taccgtcagc taacaggtgg gtaccgaagt tcaaatcgaa
     1741 gaaaactgag aaaaaacttg cccctgatgg caaaacggaa ctgctcgata aagatgaagt
     1801 ggagcgcaag atgaagtcac ttttgaacaa gttgactctg gagatgtttg atgccattag
     1861 ttctgaaatc ctggcaatcg ccaacatcag cgtatgggag acgaacggcg agacgcttaa
     1921 ggcggtaata gaacaaatct ttctcaaagc gtgtgacgag ccgcattgga gctccatgta
     1981 tgctcagctg tgcggcaaag tagtgaaaga actcaatcca gacataacag atgaaaccaa
     2041 cgagggaaag actggtccga agttggtctt gcactacctg gtagctcgct gtcatgctga
     2101 attcgataaa ggttggactg ataagcttcc gacgaatgaa gacggaaccc cgcttgaacc
     2161 cgagatgatg tctgaggagt actatgctgc cgcgtccgcc aagaggcgcg gtctgggact
     2221 tgtccgcttt attggcttcc tctatcgtct taaccttctt acaggcaaaa tgatgtttga
     2281 atgttttagg agactgatga aggacctcac tgatagtcct agtgaggaga cgctggaatc
     2341 cgttgttgag ctcctgaata ccgtaggaga gcagttcgag actgactctt tcaggaccgg
     2401 acaggcgact cttgaaggct cgcagttgct cgactcgttg tttggaatct tggataatat
     2461 tatacaaaca gccaagataa gcagtagaat aaaatttaaa cttatagata ttaaggagct
     2521 caggcacgac aaaaattgga atagtgacaa aaaagataac ggaccgaaaa ctatacaaca
     2581 gattcacgag gaagaagaga ggcaaaggca gcttaagaac aactctagga gcaactcaag
     2641 acgcacaaac aattcgtcca accgtcacag cttccgcaga gacgctccgc cagcctcgaa
     2701 ggattcgttt ataaccacaa gaacttatag ccagcgcaac agccaaagag cgccaccgcc
     2761 aaaggaagaa ccagctgccc caacaagcac agctacgaac atgttctctg cgctcatggg
     2821 tgagtccgat gatgaagaag gcgcgccgta atagaagctt gtcgagaagt actagaggat
     2881 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2941 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3001 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3061 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg
     3121 atcactgctt gagcctagga gatccgaacc agataagtga aagcctgcat ccgcagatct
     3181 taattaataa aacacgatac attgttatta gtacatttat taagcgctag attctgtgcg
     3241 ttgttgattt acagacaatt gttgtacgta ttttaataat tcattaaatt tataatcttt
     3301 agggtggtat gttagagcga aaatcaaatg attttcagcg tctttatatc tgaatttaaa
     3361 tattaaatcc tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca agggttgttt
     3421 ttccgaaccg atggctggac tatctaatgg attttcgctc aacgccacaa aacttgccaa
     3481 atcttgtagc agcaatctag ctttgtcgat attcgtttgt gttttgtttt gtaataaagg
     3541 ttcgacgtcg ttcaaaatat tatgcgcttt tgtatttctt tcatcactgt cgttagtgta
     3601 caattgactc gacgtaaaca cgttaaataa cgcttggaca tatttaacat cgggcgtgtt
     3661 agctttatta ggccgattat cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga
     3721 agacgatttt gccatagcca cacgacgcct attaattgtg tcggctaaca cgtccgcgat
     3781 caaatttgta gttgagcttt ttgggcctca tgggccttcc gctcactgcc cgctttccag
     3841 tcgggaaacc tgtcgtgcca gctgcattaa catggtcata gctgtttcct tgcgtattgg
     3901 gcgctctccg cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg
     3961 gtgcctaatg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4441 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4741 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4801 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4861 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4921 cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4981 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5041 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5101 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5161 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5221 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5281 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5341 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5401 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5461 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5521 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5581 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5641 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5701 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5761 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5821 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5881 agaatagacc gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc
     5941 tgcgcaactg ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg
     6001 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     6061 cgttgtaaaa cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattggc
     6121 ggaaggccgt caaggccgtg acgttaaaac tattaagcca tccaatcgac cgttagtcga
     6181 atcaggaccg ctggtgcgag aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga
     6241 gtccgctcat tagagcgtca tgtttagaca agaaagctac atatttaatt gatcccgatg
     6301 attttattga taaattgacc ctaactccat acacggtatt ctacaatggc ggggttttgg
     6361 tcaaaatttc cggactgcga ttgtacatgc tgttaacggc tccgcccact attaatgaaa
     6421 ttaaaaattc caattttaaa aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag
     6481 aaggaaagaa aaatgtcgtg gacatgctga acaacaagat taatatgcct ccgtgtataa
     6541 aaaaaatatt gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga
     6601 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
     6661 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
     6721 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
     6781 ctgtggacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
     6841 attgaataat aaaacaatta taaatgctaa atttgttttt tattaacgat acaaaccaaa
     6901 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
     6961 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
     7021 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
     7081 ctcctcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
     7141 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
     7201 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
     7261 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
     7321 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
     7381 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
     7441 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
     7501 cacaaactaa tatcaca
//
