LOCUS       sequence-291082-        6505 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 6505)
  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 6505)
  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-del725-755-del790-812_AE
FEATURES             Location/Qualifiers
     source          1..6505
                     /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    3293..3414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3330..3349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3384..3403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3421..3876)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3579)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3770..3789
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3903..4007
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4009..4366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4009..4200
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4032..4052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4217..4352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4401..5195
                     /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             4401..5195
                     /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(4455..4474)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5065..5084
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5383..5402)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5427..5474
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5803..6391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6292..6311
                     /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 ctatgcatta tatgggctct cctaggggat atctggatcc
     2401 cagcttggga ctttcgccag gtggtggtct cccgccgtcg catcacagtc ccattaccag
     2461 gctcatagtg ccgagccgct acgattcgcg ctttagcggg tttggcggtg gcacgcccag
     2521 gattccttcg cccagagaat accaagccaa tccggtagcg ccaacgcaac gcaatgccaa
     2581 cccaccgaat tttagtgtga acagtaacct gcacaagggc tacatgttgc ccgccagcgg
     2641 tggagatgtc ttccatctgg ccaatccgtg ggagcaagca tacctggcac agcagcaaca
     2701 tcctccacag catccccaac agcagcagcc accgtccagc aagccgaatc ccagccgacc
     2761 gttgtcgatt ttacccgcaa cagctgatac ctcatttttt gaaaagaaac cgccgaatag
     2821 tgttagcata gatctggaca gggtcccgga ggtcaatgtg gatgcagtgc ctctgttccg
     2881 gcgatccaac aataaccaca ataacaacaa tagtaactcc cacaataaca acaacaacaa
     2941 tcatggctca tcactgctgt tctggaacaa tacgggtaaa gattcagcca actttgtacg
     3001 atcagactcc atattggatg acgatgcgtc cacattctat ggcggcttca agagtgacag
     3061 aaacgatagc ttcaatgcca ataagcagca acagccgatg tggggcagga aaccgcaact
     3121 gatgtcggaa gacagtttcg acagtgggat caaggtggac tagggatcca ccggatctag
     3181 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3241 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3301 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3361 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3421 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3481 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3541 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3601 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3661 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3721 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3781 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3841 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3901 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3961 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4021 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4081 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4141 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4201 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4261 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4321 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4381 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4441 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4501 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4561 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4621 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4681 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4741 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4801 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4861 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4921 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4981 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5041 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5101 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5161 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5221 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5281 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5341 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5401 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5461 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5521 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5581 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5641 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5701 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5761 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5821 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5881 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5941 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6001 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6061 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6361 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6421 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6481 ggataaccgt attaccgcca tgcat
//
