LOCUS       sequence-291058-        8327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin-ILLVtoEEEE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8327)
  AUTHORS   Sgromo A, Raisch T, Bawankar P, Bhandari D, Chen Y, Kuzuoglu-Ozturk 
            D, Weichenrieder O, Izaurralde E
  TITLE     A CAF40-binding motif facilitates recruitment of the CCR4-NOT 
            complex to mRNAs targeted by Drosophila Roquin.
  JOURNAL   Nat Commun. 2017 Feb 6;8:14307. doi: 10.1038/ncomms14307.
  PUBMED    28165457
REFERENCE   2  (bases 1 to 8327)
  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: "2017-02-6"; volume: "8:14307. doi"; pages: " 
            10.1038/ncomms14307"
COMMENT     Sequence Label: pcDNA3.1-MS2-HA-DmRoquin-ILLVtoEEEE_AE
FEATURES             Location/Qualifiers
     source          1..8327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3883..3901)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3884..3901)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3921..3938)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3927..4151
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4197..4625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4284..4303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4494..4515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4634..4654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4639..4968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4819..4954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4881..4900
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5035..5829
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5089..5108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5699..5718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6003..6124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6040..6059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6094..6113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6173..6189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6173..6189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6186..6208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6197..6213
                     /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)."
     promoter        complement(join(6221..6227,6228..6245,6246..6251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6266..6287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6404..6421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6575..7160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6655..6674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7331..8122,8123..8191))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7954..7973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8192..8296)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccgaa
     1381 ttcccgggtc gactcgagtc tagagcggcc gccatgccga ttcaggctcc ctcatggaca
     1441 gattttctga actgtcccat ctgctgcaat gaattcgcgg ccagccagcg atgtccggtc
     1501 agtctgggct gcgggcacac catctgcaag ctatgcctga cgaccctcta caaccggcag
     1561 tgtcccttcg atcagaccgt gattgtcagt gacatcgaca atttgcccat caaccatgcc
     1621 ttgctccagt tggtcaagga ctcggagctc ctcgagctgg caccaccacc gcccagtgtg
     1681 cagaagctgg agccggagca ccttaagtgt taccaactgg gtcagcggtg catcgaggag
     1741 ctggctttgc acttgaagtc cttcctcaac ctgaacggaa atggcaatct tctaactcga
     1801 cccatgttac ggaaactggt tactctggtc aactgccagc tgatggagga ggagggacgg
     1861 gtgagagcgc ttcgagcagc cagatcccta ggcgaacgca cagtcaccga gctgatcctg
     1921 cagcaccaaa atccccagca gttgagctca aatctgtggg ctgcagtgcg gacaagagga
     1981 tgccagttcc ttggacccgc catgcaggag gaggtgctca aactggtgct gctggccttg
     2041 gaggagggat cagcgctttc gcgcaaagtg ctggtcatgt tcgtcgtgca gcgactagag
     2101 ccgaagtttc cccaggcctc caagacgagt atcggacatg tggtacagct gctctaccgc
     2161 gccagctgct tcaaggtgtc caagcgggag gcggactcgt cgttgatgca gctcaaagag
     2221 gaattccgta cgtatgacgc cctacgccgg gagcacgatg cccaaatcgt tcagatagcc
     2281 accgaggcgg gtctgcgaat tgcaccggaa caatggtctt ccctattgta tggcgacgtg
     2341 atgcataaaa gccacatgca gagcatcatc gataagctgc agacgccaag ctcctttgcg
     2401 caatcagtcc aggaattggt catcgccttg cagcggacca gcgatcctgc gaagctatcc
     2461 agtttgcatc atcatctgaa gtacctggcc aatattgatc cctgtgcgga ggttgccccg
     2521 tggtcggttt tggctgaggc cttggatgca gtgcggcatt cggtcgttgg actggtgaac
     2581 ttccttcagc accacggagt gcgcaaggcg caggacggca tcagtggcgg tggcagtggt
     2641 gggacgacca atagcaatcc gaagtacaag atcagcttat gcagggactt gaatgtgcgg
     2701 cgggtgtgtc cccgaggttc tagctgtacc tttgcccatt cccaggagga ggtagaacgc
     2761 tatcgtgcac ggaatcgagg aaaacatatg aagacaccgt tggcgctgca gggacctcca
     2821 gcggtaggcg taggagccat aaagaaacca ctaggcgagc aggagggacc gcctctaggt
     2881 aacatgccac ccatgttacc tatgtcgcct atgcattata tgggctctcc taggggatat
     2941 ctggatccca gcttgggact ttcgccaggt ggtggtctcc cgccgtcgca tcacagtccc
     3001 attaccaggc tcatagtgcc gagccgctac gattcgcgct ttagcgggtt tggcggtggc
     3061 acgcccagga ttccttcgcc cagagaatac caagccaatc cggtagcgcc aacgcaacgc
     3121 aatgccaacc caccgaattt tagtgtgaac agtaacctgc acaagggcta catgttgccc
     3181 gccagcggtg gagatgtctt ccatctggcc aatccgtggg agcaagcata cctggcacag
     3241 cagcaacatc ctccacagca tccccaacag cagcagccac cgtccagcaa gccgaatccc
     3301 agccgaccgt tgtcgatttt acccgcaaca gctgatacct cattttttga aaagaaaccg
     3361 ccgaatagtg ttagcataga tctggacagg gtcccggagg tcaatgtgga tgcagtgcct
     3421 ctgttccggc gatccaacaa taaccacaat aacaacaata gtaactccca caataacaac
     3481 aacaacaatc atggctcatc actgctgttc tggaacaata cgggtaaaga ttcagccaac
     3541 tttgtacgat cagactccat attggatgac gatgcgtcca cattcgatgt gcccactggc
     3601 tcctccatgc tgagtatcta cggcccgatt tgccccaaga gcagtacgac cggcaactgg
     3661 aacaactttg atctgggata tggcggcttc aagagtgaca gaaacgatag cttcaatgcc
     3721 aataagcagc aacagccgat gtggggcagg aaaccgcaac tgatgtcgga agacagtttc
     3781 gagggcggcg aagatagtgg aatgatgctg caggaagaga agaatgaagt ggacatcgaa
     3841 gatcccgatg acagtgggat caaggtggac taggggccct attctatagt gtcacctaaa
     3901 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3961 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4021 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4081 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4141 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     4201 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4261 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4321 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4381 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4441 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4501 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4561 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4621 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4741 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4801 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4861 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4921 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4981 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5041 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5101 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5161 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5221 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5281 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5341 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5401 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5461 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5521 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5581 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5641 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5701 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5761 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5821 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5881 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5941 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6001 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6061 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6121 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6181 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6241 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6301 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6361 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6421 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6481 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6541 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6601 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6661 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6721 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6781 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6841 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6901 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6961 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7021 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7081 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     7141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7261 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7321 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7381 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7441 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7501 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7561 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7621 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7681 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7741 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7801 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7861 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7921 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7981 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8041 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8101 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8161 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8221 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8281 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
