LOCUS       sequence-290049-        8969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of Dm4E-T-del312-356.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8969)
  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 8969)
  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-del312-356_T
FEATURES             Location/Qualifiers
     source          1..8969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /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(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6254..6295
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6305..6322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6345..6362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6395..6529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6419..6438)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6473..6492
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7151..7739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7231..7250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7910..8701,8702..8770))
                     /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     8533..8552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8771..8875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8943..8961
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttgaattcta tggatacatc aaagattagt gccaggtact cgaaagtaga
     3361 cctgttagct ctaagatatg aaggtaaaag tcgacagcgc ccgcaatgtt caacacgact
     3421 tgaattgcag acgctaggtt tttggaaaat taatctgaac acagctgcgt taaccgtgtc
     3481 cagcgcctat tctaatcaaa acaagaatcg actttctcca gaagcagata actcgagctt
     3541 aatttgttcc aatagtagct caataagttc acgtcgcgct atgagaaatc gagaacgggc
     3601 aaacaactat taccagcgct tcgtacctac cgattctctt ctaataagtg gggaggataa
     3661 agataaggat gcgctttctc atggacagcc ctataagtcg aatatcatcg atcaccgtag
     3721 catatcatct tcacatttaa tgccggcatt tgcaaagagg cgttttgtca tatcgaaagg
     3781 cagcaacagc gaggaaagca acgaggggat aaacacttgt gcaagcaagg gcaaggcagc
     3841 cagttcgcct agccgtaagg gtagcgagtt ggatactgct gagacatgtt tgaattttgt
     3901 tcagccggat cacgaccaat gcatgtcatc ttcacctaca ttttcaacat cccgccaaga
     3961 aaggcggatt ggtagcggtc ggctgttgcc aagaagtgat aattgggatt acaaaaacga
     4021 aaagaccgta gaagccagca ttgagaatga aaaggaaaca tctccaaatg gatctggaag
     4081 tacaagcagt cttaatcagc acaatcaaag ccaacatcgt tctcgaacat tcagtggaag
     4141 gttagttgaa cgtgttcccg aagtcacaga tcgtcgtttt caatatgact ccaaaaagtc
     4201 ttttgatcgt cagggaatta ataatcgacg catttctggt aaggagcctt tccgatccgt
     4261 aactgaagat aaaaataatc aaattcaaca acttgataaa aatttagacg cgcaagcaag
     4321 caaagatgaa gcttcgatga gaaacagcaa tgatagttta aatttccgcg aagtcattcc
     4381 aagtgatgaa aagaagcaca ctgacgaaaa tgtagtgact tctattcaaa actcaacaga
     4441 tctcggccat cccaataaaa acaaaccaat tcagatgcag cccagtcaga atcctgaaag
     4501 tgaatttaat tttgatgcct ttcttaatat gcatcccttg gataattccg ttttgagtaa
     4561 tgatgaaact ggaaaaagtg actcaaaagg aacatcccgt tttagccgat ggtttcgaca
     4621 aaaagaagct gcaaataata atgagtttcc agggtttcgg gaatcgcatg ctcaagaaaa
     4681 acgtggtatt ccaagtgtta aggatctgga agctcaaatg ataaaagtgg atatgaggac
     4741 tgatttaata aatccaatag cagggtcatt atgtcagaca gtgcaaatgg aaaaacctat
     4801 tgctagagac actgaggcat tcaaaaaact tttgcaacag ttgggttctc aagccaggca
     4861 gcatcatcca tgcaacgacg attgtcgtac aataaacctc tcaaatatag caaatcatgt
     4921 acaccttgag tcaaaattac atcaaaaaat aaatgatggt catttgcagc aaccagagtt
     4981 aagtgttaac gtgccaacca tgccaactag tagtcatgta tttttgcaaa agcgtttgga
     5041 aatccaacat ttaattcaaa gacttcattg tggagacgtt tcacatgatt ttttggaaaa
     5101 agaattggat aatcccagca ccccagcagc cacaaaggat gtcattgcca cagtgttgaa
     5161 tgaatattct cattctaaaa gaaatccggt ggtcacaggc gatcccaata tttttaccca
     5221 acaatcgttt cttcagcctc aatcagtcca tcagcactac tctcaagaat tgcattcgca
     5281 aaatactgct aaccatacga ttaatcagct aatttcccac ggtaactctc caactccgtt
     5341 ggcatttact cctacttcag ttttaagaaa aatgacagcc gataaggata cgcaatcacc
     5401 aagcacttat tgtcagaacc cacaatatca tgtacaccaa caaaatgcca agcaagtcgg
     5461 tactcgtgag aacgtacttg agccacagct gaccgcaaca atggctgtac agccccgaat
     5521 gatattaggc ggtggaaact ttgccattgg ccaaaacaat caacatttgt ctcccaacat
     5581 gtcacaaagt cgtaatcaac aagtgttaaa gtggacttct ggcaatatgc aaatggtcca
     5641 cggaaaaaca tttggccgac ctattctgaa aggaggtctt aattcaatgc cgcattccaa
     5701 ttctgcactt ccttttactg cccataaaat tgaaatgcaa cccattcatc aaccacatct
     5761 acagcagcag caacatagat ttaaagcagt tcaatctgtg gaatcaaatc tgaataccga
     5821 aagtgttcac caaaacatta catctcctgt tggatggcac caactttaca tgcaacatca
     5881 gcaacaacat catcacacac gacagcagct tagtcaaaga gtgatttatg gagaaatgca
     5941 ccgtcaatca aatccacaaa tgtctccgcc agtcccaggt ttttctgaca gctccgattc
     6001 cggtaatgtg atcaaagcaa acagcctcac atctcctagc taccagagag atgagaggat
     6061 ttcatcacca accaatcaat tggcgcaatg gttttcccct gagcttcttg caaaagcctc
     6121 tgctggaaaa ctgccacttt taaatgtgaa ccaagcactc agtttggaag aatttgaacg
     6181 aagcattcaa cattcctcag gcgtcgtgca taactaaggc ggccgctcga gtctagaggg
     6241 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     6301 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     6361 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     6421 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     6481 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     6541 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     6601 aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc ttgagagcct
     6661 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     6721 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     6781 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     6841 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     6901 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
     6961 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
     7021 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
     7081 aaggatcgct cgcggctctt accagccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7321 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     7381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7561 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7801 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7861 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7981 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8041 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8101 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8161 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8221 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8281 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8341 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8401 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8461 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8521 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8581 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8641 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8701 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8761 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8881 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     8941 aaaataggcg tatcacgagg ccctttcgt
//
