LOCUS       sequence-357213-        9080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of Dm4E-T-L36AL39A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9080)
  AUTHORS   Igreja C, Peter D, Weiler C, Izaurralde E
  TITLE     4E-BPs require non-canonical 4E-binding motifs and a lateral surface
            of eIF4E to repress translation.
  JOURNAL   Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.
  PUBMED    25179781
REFERENCE   2  (bases 1 to 9080)
  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"; journalName: "Nat 
            Commun."; date: "2014-09-2"; volume: "5:4790. doi"; pages: " 
            10.1038/ncomms5790"
COMMENT     Sequence Label: pAc5.1B-EGFP-Dm4E-T-L36AL39A_T
FEATURES             Location/Qualifiers
     source          1..9080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2630..2651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2891..2910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3238..3259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6386..6427
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6437..6454
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6477..6494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6527..6661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6551..6570)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6605..6624
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7262..7850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7342..7361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8021..8812,8813..8881))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8644..8663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8882..8986)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9054..9072
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3061 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gccaagcttg aattctatgg atacatcaaa gattagtgcc aggtactcga aagtagacct
     3361 gttagctcta agatatgaag gtaaaagtcg acagcgcccg caatgttcaa cacgacttga
     3421 agcgcagacg gcgggttttt ggaaaattaa tctgaacaca gctgcgttaa ccgtgtccag
     3481 cgcctattct aatcaaaaca agaatcgact ttctccagaa gcagataact cgagcttaat
     3541 ttgttccaat agtagctcaa taagttcacg tcgcgctatg agaaatcgag aacgggcaaa
     3601 caactattac cagcgcttcg tacctaccga ttctcttcta ataagtgggg aggataaaga
     3661 taaggatgcg ctttctcatg gacagcccta taagtcgaat atcatcgatc accgtagcat
     3721 atcatcttca catttaatgc cggcatttgc aaagaggcgt tttgtcatat cgaaaggcag
     3781 caacagcgag gaaagcaacg aggggataaa cacttgtgca agcaagggca aggcagccag
     3841 ttcgcctagc cgtaagggta gcgagttgga tactgctgag acatgtttga attttgttca
     3901 gccggatcac gaccaatgca tgtcatcttc acctacattt tcaacatccc gccaagaaag
     3961 gcggattggt agcggtcggc tgttgccaag aagtgataat tgggattaca aaaacgaaaa
     4021 gaccgtagaa gccagcattg agaatgaaaa ggaaacatct ccaaatggat ctggaagtac
     4081 aagcagtctt aatcagcaca atcagagcca acatcgttct cgaacattca gtggaaggtt
     4141 agttgaacgt gttcccgaag tcacagatcg tcgttttcaa tatgactcca aaaagtcttt
     4201 tgatcgtcag ggaattaata atcgacgcat ttctggtaag gagcctttca gtactcaaag
     4261 ccgaagcaag cgtggcaatt cctatctcat tcacgaagaa cctgaatggt tcagtgcagg
     4321 acctaaatca cagctagaaa ctattgatct acacgggttt gaagacctgg aaaaaaacga
     4381 agagcgatcc gtaactgaag ataaaaataa tcaaattcaa caacttgata aaaatttaga
     4441 cgcgcaagca agcaaagatg aagcttcgat gagaaacagc aatgatagtt taaatttccg
     4501 cgaagtcatt ccaagtgatg aaaagaagca cactgacgaa aatgtagtga cttctattca
     4561 aaactcaaca gatctcggcc atcccaataa aaacaaacca attcagatgc agcccagtca
     4621 gaatcctgaa agtgaattta attttgatgc ctttcttaat atgcatccct tggataattc
     4681 cgttttgagt aatgatgaaa ctggaaaaag tgactcaaag ggaacatccc gttttagccg
     4741 atggtttcga caaaaagaag ctgcaaataa taatgagttt ccagggtttc gggaatcgca
     4801 tgctcaagaa aaacgtggta ttccaagtgt taaggatctg gaagctcaaa tgataaaagt
     4861 ggatatgagg actgatttaa taaatccaat agcagggtca ttatgtcaga cagtgcaaat
     4921 ggaaaaacct attgctagag acactgaggc attcaaaaaa cttttgcaac agttgggttc
     4981 tcaagccaga cagcatcatc catgcaacga cgattgtcgt acaataaacc tctcaaatat
     5041 agcaaatcat gtacaccttg agtcaaaatt acatcaaaaa ataaatgatg gtcatttgca
     5101 gcaaccagag ttaagtgtta acgtgccaac catgccgact agtagtcatg tatttttgca
     5161 aaagcgtttg gaaatccaac atttaattca aagacttcgt tgtggagacg tttcacatga
     5221 ttttttggaa aaagagttgg ataatcccag caccccagca gccacaaagg atgtcattgc
     5281 cacagtgttg agtgaatatt ctcattctaa aagaaatccg gtggtcacag gcgatcccaa
     5341 tatttttacc caacaatcgt ttcttcagcc tcaatcagtc catcagcact actctcaaga
     5401 attgcattcg caaaatactg ctaaccatac gattaatcag ctaatttccc acggtaactc
     5461 tccaactccg ttggcattta ctcctacttc agttttaaga aaaatgacag ccgataagga
     5521 tacgcaatca ccaagcactt attgtcagaa cccacaatat catgtacacc aacaaaatgc
     5581 caagcaagtc ggtactcgtg agaacgtact tgagccacag ctgaccgcaa caatggctgt
     5641 acagccccga atgatattag gcggtggaaa ctttgccatt ggccaaaaca atcaacattt
     5701 gtctcccaac atgtcacaaa gtcgtaatca acaagtgtta aagtggactt ctggcaatat
     5761 gcaaatggtc cacggaaaaa catttggccg acctattctg aaaggaggtc ttaattcaat
     5821 gccgcattcc aattctgcgc ttccttttac tgcccataaa attgaaatgc aacccattca
     5881 tcaaccacat ctacagcagc agcaacatag atttaaagca gttcaatctg tggaatcaaa
     5941 tctgaatacc gaaagtgttc accaaaacat tacatctcct gttggatggc accaacttta
     6001 catgcaacat cagcaacaac atcatcacac acgacagcag cttagtcaaa gagtgattta
     6061 tggagaaatg caccgtcaat caaatccaca aatgtctccg ccagtcccag gtttttctga
     6121 cagctccgat tccggtaatg tgatcaaagc taacagcctc acatctccta gctaccagag
     6181 agatgagagg atttcatcac caaccaatca attggcgcaa tggttttccc ctgagcttct
     6241 tgcaaaagcc tctgctggaa aactgccact tttaaatgtg aaccaagcac tcagcttgga
     6301 agaatttgaa cgaagcattc aacattcctc aggcgtcgtg cataactaag gcggccgctc
     6361 gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     6421 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     6481 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     6541 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     6601 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     6661 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     6721 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgaccgatgc
     6781 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     6841 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     6901 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     6961 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     7021 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     7081 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     7141 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     7201 agggacagct tcaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7681 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7921 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7981 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8041 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8101 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8161 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8221 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8281 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     8341 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8401 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8461 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8521 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8581 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     8641 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8701 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8761 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8821 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8881 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8941 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9001 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9061 gtatcacgag gccctttcgt
//
