LOCUS       sequence-356597-        7127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmMextli_577-620-I583RS590R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7127)
  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 7127)
  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: pnEA-NpM-DmMextli_577-620-I583RS590R-GB1_AB
FEATURES             Location/Qualifiers
     source          1..7127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             88..1188
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="METEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             1204..1227
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1629..1647)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1643..1690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1865..1870,1871..1887,1888..1893))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1920..1938)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2006..2110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2111..2179,2180..2971)
                     /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     complement(2329..2348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3142..3730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3631..3650
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3884..3901
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3916..4055
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3981..4000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4141..4163
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4157..4348)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6703..6722
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6742..6819)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7025..7044
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
       61 tgtttaactt taagaaggag atataccatg gaaactgaag aaggtaaact ggtaatctgg
      121 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
      181 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
      241 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
      301 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
      361 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
      421 gaagcgttat cgctgattta taacaaagac ctgctgccga acccgccaaa aacctgggaa
      481 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgccct gatgttcaac
      541 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
      601 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
      661 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
      721 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
      781 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
      841 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
      901 ccgaacaaag agctggcgaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
      961 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     1021 ttggcgaaag atccacgtat tgccgccacg atggaaaacg cccagaaagg tgaaatcatg
     1081 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     1141 gccagcggtc gtcagactgt cgatgaagcc cttaaagacg cgcagactgg taccggcagc
     1201 ggcctggaag ttctgttcca ggggccccat atgctcgagt ctcgcgtttc ttacgatcgt
     1261 gaacacttgc tctactaccg tatgagtccg cactcgtgga ccctgcccac cgactggcaa
     1321 aagatgcagg agacagcgcc ctcgattctg cgcaacaagg atctgcagga tgctagcatg
     1381 cagtataaac tgattctgaa tggcaaaacc ctgaaaggtg aaaccaccac cgaagcagtt
     1441 gatgcagcca ccgcagaaaa agtttttaaa cagtatgcca atgataacgg cgttgatggt
     1501 gaatggacct atgatgatgc aaccaaaacc tataccgtta ccgaataagg atcctaatag
     1561 tctagagcta gccctaggag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1621 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1681 gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc
     1741 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     1801 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     1861 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
     1921 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1981 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2041 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2101 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2161 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2221 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2281 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2341 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2401 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2461 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2521 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2581 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2641 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2701 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2761 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2821 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2881 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2941 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3001 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3061 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3121 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3181 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3241 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3301 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3361 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3421 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3481 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3541 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3601 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3661 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3721 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3781 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3841 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3901 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3961 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4021 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4081 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4141 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4201 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4261 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4321 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4381 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4441 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4501 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4561 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4621 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4681 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4741 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4801 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     4861 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     4921 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     4981 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     5041 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     5101 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     5161 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     5221 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     5281 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     5341 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     5401 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     5461 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5521 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     5581 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     5641 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaactgtc gtgccagctg
     5701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     5761 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     5821 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     5881 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     5941 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     6001 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     6061 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     6121 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     6181 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     6241 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     6301 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     6361 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     6421 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     6481 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     6541 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     6601 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     6661 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     6721 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     6781 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     6841 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     6901 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     6961 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     7021 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     7081 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaat
//
