LOCUS       sequence-289548-        9232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmCup-del580-600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9232)
  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 9232)
  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-del580-600_O
FEATURES             Location/Qualifiers
     source          1..9232
                     /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             6517..6558
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6568..6585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6608..6625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6658..6792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6682..6701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6736..6755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7414..8002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7494..7513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8173..8964,8965..9033))
                     /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     8796..8815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9034..9138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9206..9224
                     /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 ctatacccgc tcccgcctga tggacattcg
     4381 caacgggatg ttcaatgccc tgatgcacag gtccaaggag agctttgtga tgcctcgcat
     4441 cgccacctgc gacgacattg agttggaggg aaggcttcgc 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 gaagcccagc aaatacatgc ccaaacacga
     5041 cactaacaat aatcataacg tggagaacat gaataatgtt atggctaccg agcatcagca
     5101 acaaaaggaa gagaaacgcc caggcagtgg ccgttcgttt cagtttgata agttcaatca
     5161 aagccaacag aattacgaga gtagcagtta tgttaatcac caacagccgc cgcaaacgca
     5221 gcctcagcaa atgcagcagc aatcgaacac aaacacgaac aactctaaat tcatgtcatt
     5281 ctttgccaat gaggggaact cctcgtcttc ttcgctgaac gagtttttca agcaggccat
     5341 taaccagggt catggcaaca atccggagca gcccaaatcg ttgggccaca tcggacagat
     5401 gccttcggtg gatcagctgg aggccaaatg gcgtcgaaac tcactgaaca atgttggcga
     5461 gactgccaac aagcagactg ataacttcca gaagctaatt gggtctcttt cttcggctaa
     5521 gccacagtca caggcggttg gctatgatgc catctccaac tttataatgc agcaacagca
     5581 gtaccagcag cagcaacaga agcagcacct catcatacag cagcagcagc aacagaccgc
     5641 ctttttggcg agcctgcagc tgaaggcgat cctcggccgg gccgacaccc aattgctact
     5701 gctccgtctg actaaagggg aaatatccaa gcacggattg ctggtgcagt tggccaaccc
     5761 acgcttgacg gacatggatc gcgaggccat cactgctgta ttgcagttca ccaacaccca
     5821 gcagcagcag cagcagcaca agcagcagct ggacatgctc tcaagcacgg tcatcgccag
     5881 ccagttgcag aatctccaca acttggccat tgtccagcag actcttgcag ccaggcagca
     5941 gccgcagcat aatccacaga cgcaggcgcc gcaccaactg tcgcaggagg acttgcaagc
     6001 gcatgccaat gtgataatgc gaaacgccgt aatgaagcga aaaattgagg agcagacctc
     6061 gaagcttatc aatggcggtg ccaagcatca ggcacagcag caatatctaa accgtggcca
     6121 gcaacgccag gctcgaccag atgccaactc gaatgccttg ctccatgcct tgatctctgg
     6181 cggtggcaat aaccacgcct ctggttatcc tatgaaagga cagccgcagc agcatcattc
     6241 caatttgaga ttcggtgata accaaaactt ccaatctttt gagtccaacc agccgcactt
     6301 cgccacgcag tacaagcagc aataccaaca gtctcagcag caacatcccc atcagcagcc
     6361 ccaacagctg aattcgcttc atcagaacaa cgctggagct gtcaatagct tcaataaggc
     6421 ccaaatgcaa gctcaatcag ctatttccat gctgcccaac agcggggatg agtttcatta
     6481 agcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     6541 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
     6601 tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat
     6661 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     6721 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     6781 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     6841 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag
     6901 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     6961 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     7021 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     7081 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     7141 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     7201 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     7261 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     7321 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc agcaaaaggc
     7381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     7621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8461 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     8521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8761 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     8821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9181 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gt
//
