LOCUS       sequence-148418-        8013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged DmMextli in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8013)
  AUTHORS   Peter D, Weber R, Kone C, Chung MY, Ebertsch L, Truffault V, 
            Weichenrieder O, Igreja C, Izaurralde E
  TITLE     Mextli proteins use both canonical bipartite and novel tripartite 
            binding modes to form eIF4E complexes that display differential 
            sensitivity to 4E-BP regulation.
  JOURNAL   Genes Dev. 2015 Sep 1;29(17):1835-49. doi: 10.1101/gad.269068.115. 
            Epub 2015 Aug 20.
  PUBMED    26294658
REFERENCE   2  (bases 1 to 8013)
  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"; doi: 
            "10.1101/gad.269068.115"; journalName: "Genes Dev"; date: 
            "2015-09-1- 1"; volume: "29"; issue: "17"; pages: "1835-49"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmMextli
FEATURES             Location/Qualifiers
     source          1..8013
                     /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             5298..5339
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5349..5366
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5389..5406)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5439..5573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5463..5482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5517..5536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6195..6783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6275..6294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6954..7745,7746..7814))
                     /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     7577..7596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7815..7919)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7987..8005
                     /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 tggctcacac acacctggga cgcgccgtga aaaacattga
     3361 ggcgccacgc ccgttgaaga cacaatcgcg atcgtcgttg aagaacagct acctggtgat
     3421 cgaggagcta atccagttga tagacaacgt gaccgtcggt cttcagtcgt gcaacaccac
     3481 gccggaatcg atcacgctgc tcctgcacaa cctccgggtg catggaccgc aactggaggc
     3541 ggtgtccaag gatacactgg accgggcctt cgtcgttttc cgcaacgctt cgcaggacga
     3601 acggctgaat ataacgacgc gcctcaagct gctcgaactc atcgagctgc gcgccaagag
     3661 ctgggacaat gacgacacaa tcgcgtacta caagtccaag cagcagatct ccaacgtgga
     3721 gctgccttcg gaataccagt acgatgccgg cgtgcagcct ggcgccttca gcacctcccc
     3781 cacattcggc gtcagcggcg gtgttggtgg cgttaacgat ggagccgcag ccgccgctgc
     3841 cgccgtcttc aatgcagcat ctgcagctgc agccgcccaa gctgcggcca ttgcggccgt
     3901 gggcacatcc aaccagcagc acatgctgct gccgcccggt gaggtcattc gtaattcggg
     3961 caagttcccc aaaccgacca agattcctgg caagacgtac tgcaaggacg aggtcgtgat
     4021 tcgcaatgct gactccggca aagtaatggg catcaagggg cgtcgcgtgc acatgatcga
     4081 ggagctaagc gaaaccatta tatcgtttca aagagtcaac ccgggtgcca aggagcgttt
     4141 ggtgcagatt actggtccag ctgaggataa aattaactat gccaaacaac tgatggagga
     4201 caccattcgt cggaacgcct cgccggtgcg cctggaaccc gctcctgccg ttggaggatc
     4261 ctgctcctcg cttaactcgt ccaactcgga cgacgccatt gtgcaaccaa gaacgccaac
     4321 cggaagcagt ttggccaacc ggctaagctt caactcggcc caaaacttca tgaccgccac
     4381 cgctgctgca cagcagatct cgcagcagat gcaccaccaa acgcatcacc tgcagcacca
     4441 acagcagcag caggtcgccg cagtagccgc cgcagcagca gcggcggcac atgctcaggc
     4501 gacagccgct gccggtaaag tcttgcgccc caatcaacag ctgctaatgc actcatattc
     4561 cacaaacgat gcttccgtag gtgaatacaa gttcacggtc aacgtgggcc agcacctgat
     4621 caagatcacc ggcgactgct gcgaactagt gcgcgtggcc aaactcgtgc tcgatgacta
     4681 ttttagcagc tcggagtttc tggcatccat tgaggctggc gccgccttcg acggcacttc
     4741 gctggtgacc acgccatcga ctccgcttcc cggagccggg ccgccgcaat tttggctgcc
     4801 cggcacggac agtggcattg gactgaattg cgttgtttcc tcgtcagcga acaacaatgg
     4861 tgaaggcgac gacgaggtgt tcgctgagcc cagcaatgga ggatcatcga ctagcaatca
     4921 gaatggactg gccagatcgc gtcgcagcca cttttcgcgc aaggagtcca cgccggagac
     4981 caagggcgct cgggaaaagg gtgacttgga cgatttggca ggcaccaatt ccttgaaaag
     5041 caatgcatct cgcgtttctt acgatattga acacttgctc tactactcca tgagtccgca
     5101 ctcgtggacc ctgcccaccg actggcaaaa gatgcaggag acagcgccct cgattctgcg
     5161 caacaaggat ctgcaggatg agagccagcg cttcgatggt gataagtatc tggccagcat
     5221 taagacggcc gccaagcgag acattgtggc tgccgatgag gtcgagactc tggacgaata
     5281 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     5341 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     5401 cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca
     5461 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5521 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5581 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5641 agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgaccgat gcccttgaga
     5701 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     5761 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     5821 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     5881 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     5941 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     6001 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     6061 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     6121 cttcaaggat cgctcgcggc tcttaccagc cagcaaaagg ccaggaaccg taaaaaggcc
     6181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6361 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6601 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6841 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6901 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6961 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7021 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7081 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7141 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7201 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7261 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7321 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7381 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7441 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7501 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7561 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7621 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7681 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7741 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7801 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7861 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7921 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7981 tataaaaata ggcgtatcac gaggcccttt cgt
//
