LOCUS       sequence-291056-        8165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin-del725-755-del790-812.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8165)
  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 8165)
  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-del725-755-del790-812_AE
FEATURES             Location/Qualifiers
     source          1..8165
                     /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(3721..3739)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3722..3739)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3759..3776)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3765..3989
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4035..4463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4122..4141)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4332..4353
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4472..4492)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4477..4806
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4657..4792
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4719..4738
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4873..5667
                     /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(4927..4946)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5537..5556
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5841..5962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5878..5897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5932..5951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6011..6027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6011..6027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6024..6046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6035..6051
                     /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(6059..6065,6066..6083,6084..6089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6104..6125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6242..6259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6413..6998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6493..6512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7169..7960,7961..8029))
                     /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     7792..7811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8030..8134)
                     /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 cattctatgg cggcttcaag
     3601 agtgacagaa acgatagctt caatgccaat aagcagcaac agccgatgtg gggcaggaaa
     3661 ccgcaactga tgtcggaaga cagtttcgac agtgggatca aggtggacta ggggccctat
     3721 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3781 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3841 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3901 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3961 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4021 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4081 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4141 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4201 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4261 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4321 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4381 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4441 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4501 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4561 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4621 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4681 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4741 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4801 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4861 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4921 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4981 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5041 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5101 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5161 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5221 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5281 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5341 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5401 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5461 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5521 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5581 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5641 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5701 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5761 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5821 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5881 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5941 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6001 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6061 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6961 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7021 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7081 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7141 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7201 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7261 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7321 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7381 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7441 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7501 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7561 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7621 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7681 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7741 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7801 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7861 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7921 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7981 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8041 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8101 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8161 acgtc
//
