LOCUS       sequence-291055-        8234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin-del725-755.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8234)
  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 8234)
  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_AE
FEATURES             Location/Qualifiers
     source          1..8234
                     /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(3790..3808)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3791..3808)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3828..3845)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3834..4058
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4104..4532
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4191..4210)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4401..4422
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4541..4561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4546..4875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4726..4861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4788..4807
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4942..5736
                     /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(4996..5015)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5606..5625
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5910..6031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5947..5966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6001..6020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6080..6096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6080..6096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6093..6115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6104..6120
                     /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(6128..6134,6135..6152,6153..6158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6173..6194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6311..6328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6482..7067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6562..6581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7238..8029,8030..8098))
                     /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     7861..7880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8099..8203)
                     /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 cagtttcgag ggcggcatcg atagtggaat gatgctgcag
     3721 ctggagaaga atctggtgga catcgtcgat cccgatgaca gtgggatcaa ggtggactag
     3781 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     3841 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3901 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3961 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4021 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4081 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4141 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4201 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4261 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4321 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4381 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4441 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4501 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4681 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4741 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4801 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4861 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4921 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4981 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5041 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5101 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5161 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5221 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5281 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5341 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5401 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5461 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5521 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5581 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5641 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5701 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5761 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5821 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5881 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5941 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6001 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6061 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6121 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6181 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6241 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     7021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7321 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7381 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7441 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7501 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7561 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7621 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7681 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7741 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8221 tgccacctga cgtc
//
