LOCUS       sequence-290984-        5710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin_1-500.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5710)
  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 5710)
  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: pT7-V5-SBP-C1-DmRoquin_1-500_AD
FEATURES             Location/Qualifiers
     source          1..5710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    2498..2619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2535..2554)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2589..2608
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2626..3081)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2763..2784)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2975..2994
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3108..3212
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3214..3571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3214..3405
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3237..3257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3422..3557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3606..4400
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3606..4400
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3660..3679)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4270..4289
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4588..4607)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4632..4679
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5008..5596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5497..5516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctcaag cttatatgcc gattcaggct ccctcatgga cagattttct
      901 gaactgtccc atctgctgca atgaattcgc ggccagccag cgatgtccgg tcagtctggg
      961 ctgcgggcac accatctgca agctatgcct gacgaccctc tacaaccggc agtgtccctt
     1021 cgatcagacc gtgattgtca gtgacatcga caatttgccc atcaaccatg ccttgctcca
     1081 gttggtcaag gactcggagc tcctcgagct ggcaccacca ccgcccagtg tgcagaagct
     1141 ggagccggag caccttaagt gttaccaact gggtcagcgg tgcatcgagg agctggcttt
     1201 gcacttgaag tccttcctca acctgaacgg aaatggcaat cttctaactc gacccatgtt
     1261 acggaaactg gttactctgg tcaactgcca gctgatggag gaggagggac gggtgagagc
     1321 gcttcgagca gccagatccc taggcgaacg cacagtcacc gagctgatcc tgcagcacca
     1381 aaatccccag cagttgagct caaatctgtg ggctgcagtg cggacaagag gatgccagtt
     1441 ccttggaccc gccatgcagg aggaggtgct caaactggtg ctgctggcct tggaggaggg
     1501 atcagcgctt tcgcgcaaag tgctggtcat gttcgtcgtg cagcgactag agccgaagtt
     1561 tccccaggcc tccaagacga gtatcggaca tgtggtacag ctgctctacc gcgccagctg
     1621 cttcaaggtg tccaagcggg aggcggactc gtcgttgatg cagctcaaag aggaattccg
     1681 tacgtatgac gccctacgcc gggagcacga tgcccaaatc gttcagatag ccaccgaggc
     1741 gggtctgcga attgcaccgg aacaatggtc ttccctattg tatggcgacg tgatgcataa
     1801 aagccacatg cagagcatca tcgataagct gcagacgcca agctcctttg cgcaatcagt
     1861 ccaggaattg gtcatcgcct tgcagcggac cagcgatcct gcgaagctat ccagtttgca
     1921 tcatcatctg aagtacctgg ccaatattga tccctgtgcg gaggttgccc cgtggtcggt
     1981 tttggctgag gccttggatg cagtgcggca ttcggtcgtt ggactggtga acttccttca
     2041 gcaccacgga gtgcgcaagg cgcaggacgg catcagtggc ggtggcagtg gtgggacgac
     2101 caatagcaat ccgaagtaca agatcagctt atgcagggac ttgaatgtgc ggcgggtgtg
     2161 tccccgaggt tctagctgta cctttgccca ttcccaggag gaggtagaac gctatcgtgc
     2221 acggaatcga ggaaaacata tgaagacacc gttggcgctg cagggacctc cagcggtagg
     2281 cgtaggagcc ataaagaaac cactaggcga gcaggaggga ccgcctctag gtaacatgcc
     2341 acccatgtta cctatgtcgc ctatgtaggg atccaccgga tctagataac tgatcataat
     2401 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2461 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2521 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2581 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     2641 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2701 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2761 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2821 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2881 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2941 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3001 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3061 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3121 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3181 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3241 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3301 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3481 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3541 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3601 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3661 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3721 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3781 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3841 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3901 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3961 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4021 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4081 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4141 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4201 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4261 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4321 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4381 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4441 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4501 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4561 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4621 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4681 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4741 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4801 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4861 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4921 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4981 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5041 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5101 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5161 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5221 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5281 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5341 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5401 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5461 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5521 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5581 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5641 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5701 cgccatgcat
//
