LOCUS       sequence-290026-        7807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of Dm4E-T_1-58-DmCup_417-1133.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7807)
  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 7807)
  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-lambdaN-HA-Dm4E-T_1-58-DmCup_417-1133_T
FEATURES             Location/Qualifiers
     source          1..7807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2588..2653
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             2678..2704
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5092..5133
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5143..5160
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5183..5200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5233..5367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5257..5276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5311..5330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5989..6577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6069..6088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6748..7539,7540..7608))
                     /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     7371..7390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7609..7713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7781..7799
                     /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 gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac aagctcaatg
     2641 gaaagctgca aacggtggat ctcctccaca tatggcttac ccatacgatg ttccagatta
     2701 cgctcctcca agcttgaatt ctatggatac atcaaagatt agtgccaggt actcgaaagt
     2761 agacctgtta gctctaagat atgaaggtaa aagtcgacag cgcccgcaat gttcaacacg
     2821 acttgaattg cagacgctag gtttttggaa aattaatctg aacacagctg cgttaaccgt
     2881 gtccagcgcc tattctttaa ttaatatgcc tgccttcttc aaaaacaaaa ataagccgaa
     2941 cctaatcagt gacgagtcca tcatccagag ccaaccgccc cagccgcaaa cggagttcca
     3001 agatccggcc attgttaacc aaagacgcat tggcagcggt cgcttaaacc attccaagtg
     3061 gggttacaac gacgaagact accactcgta ccacaatggc aagtcgcagc atatggagga
     3121 ggtcaactcg aagaacagca aaaacatgac ggtgctgcag ttcttcgaca acggcgagat
     3181 cagcagtcag cctcaaagac ggccgaacac tcctgtgatg gggatgtcta tcaatcgatc
     3241 cgagaacgac accctgcact cgaacgagtc cagtgaagat ttgagtcgcg ccaacgagaa
     3301 ctatgtaaag cgcgtaatgt ctggattcct ggttgtatcg aagcccaagt cacgagatgt
     3361 cgaggataga catcaccggc ggtacaggaa ccagaacgag gagcctgagt ggtttagctg
     3421 tggacccacg tctagactgg acactataga gttgtgtggg ttcgatgagg acgaggagaa
     3481 aatgctaaag gagggcaaca aaaaccacgg attaggcgaa acagagcgag aaacgtcaaa
     3541 acaaaagatg gaccataagt acaaatggac gcatgcggag ccgatgggtc gaagcaaata
     3601 catgcccaaa cacgacacta acaataatca taacgtggag aacatgaata atgttatggc
     3661 taccgagcat cagcaacaaa aggaagagaa acgcccaggc agtggccgtt cgtttcagtt
     3721 tgataagttc aatcaaagcc aacagaatta cgagagtagc agttatgtta atcaccaaca
     3781 gccgccgcaa acgcagcctc agcaaatgca gcagcaatcg aacacaaaca cgaacaactc
     3841 taaattcatg tcattctttg ccaatgaggg gaactcctcg tcttcttcgc tgaacgagtt
     3901 tttcaagcag gccattaacc agggtcatgg caacaatccg gagcagccca aatcgttggg
     3961 ccacatcgga cagatgcctt cggtggatca gctggaggcc aaatggcgtc gaaactcact
     4021 gaacaatgtt ggcgagactg ccaacaagca gactgataac ttccagaagc taattgggtc
     4081 tctttcttcg gctaagccac agtcacaggc ggttggctat gatgccatct ccaactttat
     4141 aatgcagcaa cagcagtacc agcagcagca acagaagcag cacctcatca tacagcagca
     4201 gcagcaacag accgcctttt tggcgagcct gcagctgaag gcgatcctcg gccgggccga
     4261 cacccaattg ctactgctcc gtctgactaa aggggaaata tccaagcacg gattgctggt
     4321 gcagttggcc aacccacgct tgacggacat ggatcgcgag gccatcactg ctgtattgca
     4381 gttcaccaac acccagcagc agcagcagca gcacaagcag cagctggaca tgctctcaag
     4441 cacggtcatc gccagccagt tgcagaatct ccacaacttg gccattgtcc agcagactct
     4501 tgcagccagg cagcagccgc agcataatcc acagacgcag gcgccgcacc aactgtcgca
     4561 ggaggacttg caagcgcatg ccaatgtgat aatgcgaaac gccgtaatga agcgaaaaat
     4621 tgaggagcag acctcgaagc ttatcaatgg cggtgccaag catcaggcac agcagcaata
     4681 tctaaaccgt ggccagcaac gccaggctcg accagatgcc aactcgaatg ccttgctcca
     4741 tgccttgatc tctggcggtg gcaataacca cgcctctggt tatcctatga aaggacagcc
     4801 gcagcagcat cattccaatt tgagattcgg tgataaccaa aacttccaat cttttgagtc
     4861 caaccagccg cacttcgcca cgcagtacaa gcagcaatac caacagtctc agcagcaaca
     4921 tccccatcag cagccccaac agctgaattc gcttcatcag aacaacgctg gagctgtcaa
     4981 tagcttcaat aaggcccaaa tgcaagctca atcagctatt tccatgctgc ccaacagcgg
     5041 ggatgagttt cattaagcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct
     5101 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat
     5161 tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc tcgctgatca
     5221 gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca aaccacaact
     5281 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5341 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5401 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     5461 gctgattatg atcagtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     5521 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     5581 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     5641 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     5701 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     5761 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     5821 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     5881 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     5941 cagccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6001 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6061 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6121 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6181 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6241 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6301 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6361 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6421 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6481 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6541 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6601 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6661 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6721 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6781 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6841 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6901 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6961 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7021 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc
     7081 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7141 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7201 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7261 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7321 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     7381 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7441 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7501 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7561 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7621 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7681 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7741 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7801 ctttcgt
//
