LOCUS       sequence-290820-        9104 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Insect Expression of Dm4E-T-F41DW42D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9104)
  AUTHORS   Peter D, Igreja C, Weber R, Wohlbold L, Weiler C, Ebertsch L, 
            Weichenrieder O, Izaurralde E
  TITLE     Molecular architecture of 4E-BP translational inhibitors bound to 
            eIF4E.
  JOURNAL   Mol Cell. 2015 Mar 19;57(6):1074-87. doi: 
            10.1016/j.molcel.2015.01.017. Epub 2015 Feb 19.
  PUBMED    25702871
REFERENCE   2  (bases 1 to 9104)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2015.01.017"; journalName: "Mol Cell"; date: 
            "2015-03-19- 19"; volume: "57"; issue: "6"; pages: "1074-87"
COMMENT     Sequence Label: pAc5.1B-EGFP-Dm4E-T-F41DW42D_AB
FEATURES             Location/Qualifiers
     source          1..9104
                     /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             6389..6430
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6440..6457
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6480..6497)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6530..6664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6554..6573)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6608..6627
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7286..7874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7366..7385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8045..8836,8837..8905))
                     /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     8668..8687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8906..9010)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9078..9096
                     /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 acgctaggtg acgacaaaat 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 acaatcagag ccaacatcgt tctcgaacat tcagtggaag
     4141 gttagttgaa cgtgttcccg aagtcacaga tcgtcgtttt caatatgact ccaaaaagtc
     4201 ttttgatcgt cagggaatta ataatcgacg catttctggt aaggagcctt tcagtactca
     4261 aagccgaagc aagcgtggca attcctatct cattcacgaa gaacctgaat ggttcagtgc
     4321 aggacctaaa tcacagctag aaactattga tctacacggg tttgaagacc tggaaaaaaa
     4381 cgaagagcga tccgtaactg aagataaaaa taatcaaatt caacaacttg ataaaaattt
     4441 agacgcgcaa gcaagcaaag atgaagcttc gatgagaaac agcaatgata gtttaaattt
     4501 ccgcgaagtc attccaagtg atgaaaagaa gcacactgac gaaaatgtag tgacttctat
     4561 tcaaaactca acagatctcg gccatcccaa taaaaacaaa ccaattcaga tgcagcccag
     4621 tcagaatcct gaaagtgaat ttaattttga tgcctttctt aatatgcatc ccttggataa
     4681 ttccgttttg agtaatgatg aaactggaaa aagtgactca aaaggaacat cccgttttag
     4741 ccgatggttt cgacaaaaag aagctgcaaa taataatgag tttccagggt ttcggggatc
     4801 gcatgctcaa gaaaaacgtg gtattccaag tgttaaggat ctggaagctc aaatgataaa
     4861 agtggatatg aggactgatt taataaatcc aatagcaggg tcattatgtc agacagtgca
     4921 aatggaaaaa cctattgcta gagacactga ggcattcaaa aaacttttgc aacagttggg
     4981 ttctcaagcc aggcagcatc atccatgcaa cgacgattgt cgtacaataa acctctcaaa
     5041 tatagcaaat catgtacacc ttgagtcaaa attacatcaa aaaataaatg atggtcattt
     5101 gcagcaacca gagttaagtg ttaacgtgcc aaccatgcca actagtagtc atgtattttt
     5161 gcaaaagcgt ttggaaatcc aacatttaat tcaaagactt cgttgtggag acgtttcaca
     5221 tgattttttg gaaaaagaat tggataatcc cagcacccca gcagccacaa aggatgtcat
     5281 tgccacagtg ttgagtgaat attctcattc taaaagaaat ccggtggtca caggcgatcc
     5341 caatattttt acccaacaat cgtttcttca gcctcaatca gtccatcagc actactctca
     5401 agaattgcat tcgcaaaata ctgctaacca tacgattaat cagctaattt cccacggtaa
     5461 ctctccaact ccgttggcat ttactcctac ttcagtttta agaaaaatga cagccgataa
     5521 ggatacgcaa tcaccaagca cttattgtca gaacccacaa tatcatgtac accaacaaaa
     5581 tgccaagcaa gtcggtactc gtgagaacgt acttgagcca cagctgaccg caacaatggc
     5641 tgtacagccc cgaatgatat taggcggtgg aaactttgcc attggccaaa acaatcaaca
     5701 tttgtctccc aacatgtcac aaagtcgtaa tcaacaagtg ttaaagtgga cttctggcaa
     5761 tatgcaaatg gtccacggaa aaacatttgg ccgacctatt ctgaaaggag gtcttaattc
     5821 aatgccgcat tccaattctg cgcttccttt tactgcccat aaaattgaaa tgcaacccat
     5881 tcatcaacca catctacagc agcagcaaca tagatttaaa gcagttcaat ctgtggaatc
     5941 aaatctgaat accgaaagtg ttcaccaaaa cattacatct cctgttggat ggcaccaact
     6001 ttacatgcaa catcagcaac aacatcatca cacacgacag cagcttagtc aaagagtgat
     6061 ttatggagaa atgcaccgtc aatcaaatcc acaaatgtct ccgccagtcc caggtttttc
     6121 tgacagctcc gattccggta atgtgatcaa agctaacggc ctcacatctc ctagctacca
     6181 gagagatgag aggatttcat caccaaccaa tcaattggcg caatggtttt cccctgagct
     6241 tcttgcaaaa gcctctgctg gaaaactgcc acttttaaat gtgaaccaag cactcagctt
     6301 ggaagaattt gaacgaagca ttcaacattc ctcaggcgtc gtgcataact aaggcggccg
     6361 ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct
     6421 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     6481 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     6541 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     6601 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     6661 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     6721 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgaccga
     6781 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     6841 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     6901 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     6961 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     7021 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
     7081 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     7141 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     7201 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag ccagcaaaag gccaggaacc
     7261 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7321 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7381 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7441 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     7501 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7561 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7621 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7681 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     7741 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7801 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7861 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7921 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7981 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8041 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8101 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8161 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8221 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8281 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8341 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8401 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8461 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8521 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8581 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8641 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     8701 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8761 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8821 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8881 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8941 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9001 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     9061 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgt
//
