LOCUS       sequence-357786-        8330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8330)
  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 8330)
  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_AD
FEATURES             Location/Qualifiers
     source          1..8330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      661..670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1099..1125
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3649..3667)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3650..3667)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3687..3704)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3693..3917
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3963..4391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4050..4069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4260..4281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4400..4420)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4405..4734
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4585..4720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4647..4666
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4801..5595
                     /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(4855..4874)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5465..5484
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5769..5890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5806..5825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5860..5879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5939..5955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5939..5955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5952..5974)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5963..5979
                     /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(5987..5993,5994..6011,6012..6017))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6032..6053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6170..6187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6341..6929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6421..6440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7100..7891,7892..7960))
                     /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     7723..7742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7961..8065)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8140..8159)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gccaccatgg cttctaactt tactcagttc gttctcgtcg acaatggcgg aactggcgac
      721 gtgactgtcg ccccaagcaa cttcgctaac ggggtcgctg aatggatcag ctctaactcg
      781 cgatcacagg cttacaaagt aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa
      841 tacaccatca aagtcgaggt gcctaaagtg gcaacccaga ctgttggtgg tgaagagctt
      901 cctgtagccg gatggagatc ttacttaaat atggaactaa ccattccaat tttcgccacg
      961 aattccgact gcgagcttat tgttaaggca atgcaaggtc tcctaaaaga tggaaacccg
     1021 attccctcgg ccatcgcggc caactccggc atctacggag gtggaggtgg atctggtgga
     1081 tctcctccac atatggctta cccatacgat gttccagatt acgctcctcc aaagcttgga
     1141 tccgaattcc cgggtcgact cgagtctaga gcggccgcca tgccgattca ggctccctca
     1201 tggacagatt ttctgaactg tcccatctgc tgcaatgaat tcgcggccag ccagcgatgt
     1261 ccggtcagtc tgggctgcgg gcacaccatc tgcaagctat gcctgacgac cctctacaac
     1321 cggcagtgtc ccttcgatca gaccgtgatt gtcagtgaca tcgacaattt gcccatcaac
     1381 catgccttgc tccagttggt caaggactcg gagctcctcg agctggcacc accaccgccc
     1441 agtgtgcaga agctggagcc ggagcacctt aagtgttacc aactgggtca gcggtgcatc
     1501 gaggagctgg ctttgcactt gaagtccttc ctcaacctga acggaaatgg caatcttcta
     1561 actcgaccca tgttacggaa actggttact ctggtcaact gccagctgat ggaggaggag
     1621 ggacgggtga gagcgcttcg agcagccaga tccctaggcg aacgcacagt caccgagctg
     1681 atcctgcagc accaaaatcc ccagcagttg agctcaaatc tgtgggctgc agtgcggaca
     1741 agaggatgcc agttccttgg acccgccatg caggaggagg tgctcaaact ggtgctgctg
     1801 gccttggagg agggatcagc gctttcgcgc aaagtgctgg tcatgttcgt cgtgcagcga
     1861 ctagagccga agtttcccca ggcctccaag acgagtatcg gacatgtggt acagctgctc
     1921 taccgcgcca gctgcttcaa ggtgtccaag cgggaggcgg actcgtcgtt gatgcagctc
     1981 aaagaggaat tccgtacgta tgacgcccta cgccgggagc acgatgccca aatcgttcag
     2041 atagccaccg aggcgggtct gcgaattgca ccggaacaat ggtcttccct attgtatggc
     2101 gacgtgatgc ataaaagcca catgcagagc atcatcgata agctgcagac gccaagctcc
     2161 tttgcgcaat cagtccagga attggtcatc gccttgcagc ggaccagcga tcctgcgaag
     2221 ctatccagtt tgcatcatca tctgaagtac ctggccaata ttgatccctg tgcggaggtt
     2281 gccccgtggt cggttttggc tgaggccttg gatgcagtgc ggcattcggt cgttggactg
     2341 gtgaacttcc ttcagcacca cggagtgcgc aaggcgcagg acggcatcag tggcggtggc
     2401 agtggtggga cgaccaatag caatccgaag tacaagatca gcttatgcag ggacttgaat
     2461 gtgcggcggg tgtgtccccg aggttctagc tgtacctttg cccattccca ggaggaggta
     2521 gaacgctatc gtgcacggaa tcgaggaaaa catatgaaga caccgttggc gctgcaggga
     2581 cctccagcgg taggcgtagg agccataaag aaaccactag gcgagcagga gggaccgcct
     2641 ctaggtaaca tgccacccat gttacctatg tcgcctatgc attatatggg ctctcctagg
     2701 ggatatctgg atcccagctt gggactttcg ccaggtggtg gtctcccgcc gtcgcatcac
     2761 agtcccatta ccaggctcat agtgccgagc cgctacgatt cgcgctttag cgggtttggc
     2821 ggtggcacgc ccaggattcc ttcgcccaga gaataccaag ccaatccggt agcgccaacg
     2881 caacgcaatg ccaacccacc gaattttagt gtgaacagta acctgcacaa gggctacatg
     2941 ttgcccgcca gcggtggaga tgtcttccat ctggccaatc cgtgggagca agcatacctg
     3001 gcacagcagc aacatcctcc acagcatccc caacagcagc agccaccgtc cagcaagccg
     3061 aatcccagcc gaccgttgtc gattttaccc gcaacagctg atacctcatt ttttgaaaag
     3121 aaaccgccga atagtgttag catagatctg gacagggtcc cggaggtcaa tgtggatgca
     3181 gtgcctctgt tccggcgatc caacaataac cacaataaca acaatagtaa ctcccacaat
     3241 aacaacaaca acaatcatgg ctcatcactg ctgttctgga acaatacggg taaagattca
     3301 gccaactttg tacgatcaga ctccatattg gatgacgatg cgtccacatt cgatgtgccc
     3361 actggctcct ccatgctgag tatctacggc ccgatttgcc ccaagagcag tacgaccggc
     3421 aactggaaca actttgatct gggatatggc ggcttcaaga gtgacagaaa cgatagcttc
     3481 aatgccaata agcagcaaca gccgatgtgg ggcaggaaac cgcaactgat gtcggaagac
     3541 agtttcgagg gcggcatcga tagtggaatg atgctgcagc tggagaagaa tctggtggac
     3601 atcgtcgatc ccgatgacag tgggatcaag gtggactagg ggccctattc tatagtgtca
     3661 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3721 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3781 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3841 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3901 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3961 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4021 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4141 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4261 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4501 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4561 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4621 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4681 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4741 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4801 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4861 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4921 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4981 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5041 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5101 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5161 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5221 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5281 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5341 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5401 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5461 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5521 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5581 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5641 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5701 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5761 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5881 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5941 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6001 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6061 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6121 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6781 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7201 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7381 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7681 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8101 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     8161 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     8221 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     8281 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
