LOCUS       sequence-290048-        9056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of Dm4E-T-del28-43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9056)
  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 9056)
  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-del28-43_T
FEATURES             Location/Qualifiers
     source          1..9056
                     /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             6341..6382
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6392..6409
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6432..6449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6482..6616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6506..6525)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6560..6579
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7238..7826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7318..7337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7997..8788,8789..8857))
                     /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     8620..8639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8858..8962)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9030..9048
                     /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 attaatctga acacagctgc
     3421 gttaaccgtg tccagcgcct attctaatca aaacaagaat cgactttctc cagaagcaga
     3481 taactcgagc ttaatttgtt ccaatagtag ctcaataagt tcacgtcgcg ctatgagaaa
     3541 tcgagaacgg gcaaacaact attaccagcg cttcgtacct accgattctc ttctaataag
     3601 tggggaggat aaagataagg atgcgctttc tcatggacag ccctataagt cgaatatcat
     3661 cgatcaccgt agcatatcat cttcacattt aatgccggca tttgcaaaga ggcgttttgt
     3721 catatcgaaa ggcagcaaca gcgaggaaag caacgagggg ataaacactt gtgcaagcaa
     3781 gggcaaggca gccagttcgc ctagccgtaa gggtagcgag ttggatactg ctgagacatg
     3841 tttgaatttt gttcagccgg atcacgacca atgcatgtca tcttcaccta cattttcaac
     3901 atcccgccaa gaaaggcgga ttggtagcgg tcggctgttg ccaagaagtg ataattggga
     3961 ttacaaaaac gaaaagaccg tagaagccag cattgagaat gaaaaggaaa catctccaaa
     4021 tggatctgga agtacaagca gtcttaatca gcacaatcaa agccaacatc gttctcgaac
     4081 attcagtgga aggttagttg aacgtgttcc cgaagtcaca gatcgtcgtt ttcaatatga
     4141 ctccaaaaag tcttttgatc gtcagggaat taataatcga cgcatttctg gtaaggagcc
     4201 tttcagtact caaagccgaa gcaagcgtgg caattcctat ctcattcacg aagaacctga
     4261 atggttcagt gcaggaccta aatcacagct agaaactatt gatctacacg ggtttgaaga
     4321 cctggaaaaa aacgaagagc gatccgtaac tgaagataaa aataatcaaa ttcaacaact
     4381 tgataaaaat ttagacgcgc aagcaagcaa agatgaagct tcgatgagaa acagcaatga
     4441 tagtttaaat ttccgcgaag tcattccaag tgatgaaaag aagcacactg acgaaaatgt
     4501 agtgacttct attcaaaact caacagatct cggccatccc aataaaaaca aaccaattca
     4561 gatgcagccc agtcagaatc ctgaaagtga atttaatttt gatgcctttc ttaatatgca
     4621 tcccttggat aattccgttt tgagtaatga tgaaactgga aaaagtgact caaaaggaac
     4681 atcccgtttt agccgatggt ttcgacaaaa agaagctgca aataataatg agtttccagg
     4741 gtttcgggaa tcgcatgctc aagaaaaacg tggtattcca agtgttaagg atctggaagc
     4801 tcaaatgata aaagtggata tgaggactga tttaataaat ccaatagcag ggtcattatg
     4861 tcagacagtg caaatggaaa aacctattgc tagagacact gaggcattca aaaaactttt
     4921 gcaacagttg ggttctcaag ccaggcagca tcatccatgc aacgacgatt gtcgtacaat
     4981 aaacctctca aatatagcaa atcatgtaca ccttgagtca aaattacatc aaaaaataaa
     5041 tgatggtcat ttgcagcaac cagagttaag tgttaacgtg ccaaccatgc caactagtag
     5101 tcatgtattt ttgcaaaagc gtttggaaat ccaacattta attcaaagac ttcattgtgg
     5161 agacgtttca catgattttt tggaaaaaga attggataat cccagcaccc cagcagccac
     5221 aaaggatgtc attgccacag tgttgaatga atattctcat tctaaaagaa atccggtggt
     5281 cacaggcgat cccaatattt ttacccaaca atcgtttctt cagcctcaat cagtccatca
     5341 gcactactct caagaattgc attcgcaaaa tactgctaac catacgatta atcagctaat
     5401 ttcccacggt aactctccaa ctccgttggc atttactcct acttcagttt taagaaaaat
     5461 gacagccgat aaggatacgc aatcaccaag cacttattgt cagaacccac aatatcatgt
     5521 acaccaacaa aatgccaagc aagtcggtac tcgtgagaac gtacttgagc cacagctgac
     5581 cgcaacaatg gctgtacagc cccgaatgat attaggcggt ggaaactttg ccattggcca
     5641 aaacaatcaa catttgtctc ccaacatgtc acaaagtcgt aatcaacaag tgttaaagtg
     5701 gacttctggc aatatgcaaa tggtccacgg aaaaacattt ggccgaccta ttctgaaagg
     5761 aggtcttaat tcaatgccgc attccaattc tgcacttcct tttactgccc ataaaattga
     5821 aatgcaaccc attcatcaac cacatctaca gcagcagcaa catagattta aagcagttca
     5881 atctgtggaa tcaaatctga ataccgaaag tgttcaccaa aacattacat ctcctgttgg
     5941 atggcaccaa ctttacatgc aacatcagca acaacatcat cacacacgac agcagcttag
     6001 tcaaagagtg atttatggag aaatgcaccg tcaatcaaat ccacaaatgt ctccgccagt
     6061 cccaggtttt tctgacagct ccgattccgg taatgtgatc aaagcaaaca gcctcacatc
     6121 tcctagctac cagagagatg agaggatttc atcaccaacc aatcaattgg cgcaatggtt
     6181 ttcccctgag cttcttgcaa aagcctctgc tggaaaactg ccacttttaa atgtgaacca
     6241 agcactcagt ttggaagaat ttgaacgaag cattcaacat tcctcaggcg tcgtgcataa
     6301 ctaaggcggc cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
     6361 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     6421 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     6481 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     6541 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     6601 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     6661 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     6721 tcagtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     6781 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     6841 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     6901 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     6961 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     7021 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     7081 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     7141 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agccagcaaa
     7201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     7441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8281 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     8341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8581 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     8641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8881 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8941 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9001 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
