LOCUS       sequence-290055-        7691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of Dm4E-T_1-738.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  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 7691)
  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_1-738_T
FEATURES             Location/Qualifiers
     source          1..7691
                     /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             4976..5017
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5027..5044
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5067..5084)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5117..5251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5141..5160)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5195..5214
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5873..6461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5953..5972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6632..7423,7424..7492))
                     /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     7255..7274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7493..7597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7665..7683
                     /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 tattctaatc aaaacaagaa tcgactttct ccagaagcag ataactcgag
     2941 cttaatttgt tccaatagta gctcaataag ttcacgtcgc gctatgagaa atcgagaacg
     3001 ggcaaacaac tattaccagc gcttcgtacc taccgattct cttctaataa gtggggagga
     3061 taaagataag gatgcgcttt ctcatggaca gccctataag tcgaatatca tcgatcaccg
     3121 tagcatatca tcttcacatt taatgccggc atttgcaaag aggcgttttg tcatatcgaa
     3181 aggcagcaac agcgaggaaa gcaacgaggg gataaacact tgtgcaagca agggcaaggc
     3241 agccagttcg cctagccgta agggtagcga gttggatact gctgagacat gtttgaattt
     3301 tgttcagccg gatcacgacc aatgcatgtc atcttcacct acattttcaa catcccgcca
     3361 agaaaggcgg attggtagcg gtcggctgtt gccaagaagt gataattggg attacaaaaa
     3421 cgaaaagacc gtagaagcca gcattgagaa tgaaaaggaa acatctccaa atggatctgg
     3481 aagtacaagc agtcttaatc agcacaatca aagccaacat cgttctcgaa cattcagtgg
     3541 aaggttagtt gaacgtgttc ccgaagtcac agatcgtcgt tttcaatatg actccaaaaa
     3601 gtcttttgat cgtcagggaa ttaataatcg acgcatttct ggtaaggagc ctttcagtac
     3661 tcaaagccga agcaagcgtg gcaattccta tctcattcac gaagaacctg aatggttcag
     3721 tgcaggacct aaatcacagc tagaaactat tgatctacac gggtttgaag acctggaaaa
     3781 aaacgaagag cgatccgtaa ctgaagataa aaataatcaa attcaacaac ttgataaaaa
     3841 tttagacgcg caagcaagca aagatgaagc ttcgatgaga aacagcaatg atagtttaaa
     3901 tttccgcgaa gtcattccaa gtgatgaaaa gaagcacact gacgaaaatg tagtgacttc
     3961 tattcaaaac tcaacagatc tcggccatcc caataaaaac aaaccaattc agatgcagcc
     4021 cagtcagaat cctgaaagtg aatttaattt tgatgccttt cttaatatgc atcccttgga
     4081 taattccgtt ttgagtaatg atgaaactgg aaaaagtgac tcaaaaggaa catcccgttt
     4141 tagccgatgg tttcgacaaa aagaagctgc aaataataat gagtttccag ggtttcggga
     4201 atcgcatgct caagaaaaac gtggtattcc aagtgttaag gatctggaag ctcaaatgat
     4261 aaaagtggat atgaggactg atttaataaa tccaatagca gggtcattat gtcagacagt
     4321 gcaaatggaa aaacctattg ctagagacac tgaggcattc aaaaaacttt tgcaacagtt
     4381 gggttctcaa gccaggcagc atcatccatg caacgacgat tgtcgtacaa taaacctctc
     4441 aaatatagca aatcatgtac accttgagtc aaaattacat caaaaaataa atgatggtca
     4501 tttgcagcaa ccagagttaa gtgttaacgt gccaaccatg ccaactagta gtcatgtatt
     4561 tttgcaaaag cgtttggaaa tccaacattt aattcaaaga cttcattgtg gagacgtttc
     4621 acatgatttt ttggaaaaag aattggataa tcccagcacc ccagcagcca caaaggatgt
     4681 cattgccaca gtgttgaatg aatattctca ttctaaaaga aatccggtgg tcacaggcga
     4741 tcccaatatt tttacccaac aatcgtttct tcagcctcaa tcagtccatc agcactactc
     4801 tcaagaattg cattcgcaaa atactgctaa ccatacgatt aatcagctaa tttcccacgg
     4861 taactctcca actccgttgg catttactcc tacttcagtt ttaagaaaaa tgacagccga
     4921 taaggatacg caatcataag gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa
     4981 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     5041 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     5101 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     5161 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     5221 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     5281 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     5341 tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     5401 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     5461 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     5521 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     5581 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     5641 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     5701 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     5761 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     5821 ttaccagcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6061 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6301 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6361 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6421 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6481 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6541 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6601 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6661 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6721 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6781 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6841 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6901 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     6961 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7021 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7081 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7141 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7201 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     7261 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7321 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7381 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7441 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7501 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7561 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7621 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     7681 ggccctttcg t
//
