LOCUS       sequence-289547-        9478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmCup-4Ala-mut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9478)
  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 9478)
  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-DmCup-4Ala-mut_O
FEATURES             Location/Qualifiers
     source          1..9478
                     /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(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6763..6804
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6814..6831
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6854..6871)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6904..7038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6928..6947)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6982..7001
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7660..8248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7740..7759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8419..9210,9211..9279))
                     /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     9042..9061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9280..9384)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9452..9470
                     /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 ttgaattctg cagatatcat ggaccctatg aaagaggagg cgatcgaatt
     3361 gaatggccat gctatgcaaa tggccgaagc tgagcaagaa aatggcgctg gcgccctgaa
     3421 aattgccacc aatgcgggtg caaccgatcg tccagcgcat cagcagcttc cgctgccagt
     3481 cgaagatcag caggatgagg tgctgacccc cgctgagaag ggcaagttcg agtacccgcc
     3541 ccctccaccg ccaccaactc cagtccaggc tccgctagcc accaaggcca ccgctctgaa
     3601 tgccagtcaa gagcatgacg acgacgaggc caactccgag aagtgggagg atccgtgcgc
     3661 tccgcccccg cctcctccgc tgccgactag cgctttcctg gccaccggac tgggctacct
     3721 caagctgccc gccttcaagc tgaaggacgc actggagaag gccatcacca agttggaggc
     3781 caacaagcgg acgctaaagg ccagccctga gtcttctcgt tccatcaaaa acaagaatgt
     3841 ggtggccctg gagatgttac ccaggcgctc aaacccagaa acgatcggcg atggatccat
     3901 gttggcatcc acatccacgg ctgttatgct gcagaccaaa aagcccgccg ttattgtgga
     3961 gatggagagg cgttgtaaga tcatcaattt actggccaaa cagaatcaaa tcctcgaatc
     4021 catttcgggg gaggccattc cgatgcatgg accctcgaag catctgcacg aggacgaggg
     4081 tctgaccctg caagttctct cggcccgagc atccacaccg tacacacagc cctccagcat
     4141 gttgtcctgc acggccgtta gttgtgacct ggagcacgat tcgccgcgga agcaagtggc
     4201 cagcaaggag gccgtgccgg agcagcagtc tagtcaggtg cagcaaaagc gtccacccag
     4261 cacaggcatt cacaagccag gaagtctgcg agcacccaag gctgttcgtc ctactactgc
     4321 tccggtggtc agcagcaagc cggtcaagag cgctacccgc tcccgcgcga tggacattcg
     4381 caacgggatg ttcaatgccc tgatgcacag gtccaaggag agctttgtga tgcctcgcat
     4441 cgccacctgc gacgacattg aggcggaggg aagggctcgc cgcatgaata tttggcgcac
     4501 ctcggacggc acaagattcc ggacccgctc caccacagct aacttaaaca tgaacaacaa
     4561 caacaacaat gagtgcatgc ctgccttctt caaaaacaaa aataagccga acctaatcag
     4621 tgacgagtcc atcatccaga gccaaccgcc ccagccgcaa acggagttcc aagatccggc
     4681 cattgttaac caaagacgca ttggcagcgg tcgcttaaac cattccaagt ggggttacaa
     4741 cgacgaagac taccactcgt accacaatgg caagtcgcag catatggagg aggtcaactc
     4801 gaagaacagc aaaaacatga cggtgctgca gttcttcgac aacggcgaga tcagcagtca
     4861 gcctcaaaga cggccgaaca ctcctgtgat ggggatgtct atcaatcgat ccgagaacga
     4921 caccctgcac tcgaacgagt ccagtgaaga tttgagtcgc gccaacgaga actatgtaaa
     4981 gcgcgtaatg tctggattcc tggttgtatc gaagcccaag tcacgagatg tcgaggatag
     5041 acatcaccgg cggtacagga accagaacga ggagcctgag tggtttagct gtggacccac
     5101 gtctagactg gacactatag agttgtgtgg gttcgatgag gacgaggaga aaatgctaaa
     5161 ggagggcaac aaaaaccacg gattaggcga aacagagcga gaaacgtcaa aacaaaagat
     5221 ggaccataag tacaaatgga cgcatgcgga gccgatgggt cgaagcaaat acatgcccaa
     5281 acacgacact aacaataatc ataacgtgga gaacatgaat aatgttatgg ctaccgagca
     5341 tcagcaacaa aaggaagaga aacgcccagg cagtggccgt tcgtttcagt ttgataagtt
     5401 caatcaaagc caacagaatt acgagagtag cagttatgtt aatcaccaac agccgccgca
     5461 aacgcagcct cagcaaatgc agcagcaatc gaacacaaac acgaacaact ctaaattcat
     5521 gtcattcttt gccaatgagg ggaactcctc gtcttcttcg ctgaacgagt ttttcaagca
     5581 ggccattaac cagggtcatg gcaacaatcc ggagcagccc aaatcgttgg gccacatcgg
     5641 acagatgcct tcggtggatc agctggaggc caaatggcgt cgaaactcac tgaacaatgt
     5701 tggcgagact gccaacaagc agactgataa cttccagaag ctaattgggt ctctttcttc
     5761 ggctaagcca cagtcacagg cggttggcta tgatgccatc tccaacttta taatgcagca
     5821 acagcagtac cagcagcagc aacagaagca gcacctcatc atacagcagc agcagcaaca
     5881 gaccgccttt ttggcgagcc tgcagctgaa ggcgatcctc ggccgggccg acacccaatt
     5941 gctactgctc cgtctgacta aaggggaaat atccaagcac ggattgctgg tgcagttggc
     6001 caacccacgc ttgacggaca tggatcgcga ggccatcact gctgtattgc agttcaccaa
     6061 cacccagcag cagcagcagc agcacaagca gcagctggac atgctctcaa gcacggtcat
     6121 cgccagccag ttgcagaatc tccacaactt ggccattgtc cagcagactc ttgcagccag
     6181 gcagcagccg cagcataatc cacagacgca ggcgccgcac caactgtcgc aggaggactt
     6241 gcaagcgcat gccaatgtga taatgcgaaa cgccgtaatg aagcgaaaaa ttgaggagca
     6301 gacctcgaag cttatcaatg gcggtgccaa gcatcaggca cagcagcaat atctaaaccg
     6361 tggccagcaa cgccaggctc gaccagatgc caactcgaat gccttgctcc atgccttgat
     6421 ctctggcggt ggcaataacc acgcctctgg ttatcctatg aaaggacagc cgcagcagca
     6481 tcattccaat ttgagattcg gtgataacca aaacttccaa tcttttgagt ccaaccagcc
     6541 gcacttcgcc acgcagtaca agcagcaata ccaacagtct cagcagcaac atccccatca
     6601 gcagccccaa cagctgaatt cgcttcatca gaacaacgct ggagctgtca atagcttcaa
     6661 taaggcccaa atgcaagctc aatcagctat ttccatgctg cccaacagcg gggatgagtt
     6721 tcattaagcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc tatccctaac
     6781 cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa
     6841 acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc agcctcgatc
     6901 gaggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     6961 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     7021 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     7081 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     7141 gatcagtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     7201 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     7261 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
     7321 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
     7381 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga
     7441 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat
     7501 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca
     7561 ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta ccagccagca
     7621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     7861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     8041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     8101 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     8161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     8221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     8281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8701 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     8761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     9001 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     9061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     9121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     9181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     9241 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9301 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9421 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgt
//
