LOCUS       sequence-291083-        6598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin-del790-812.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6598)
  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 6598)
  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-del790-812_AE
FEATURES             Location/Qualifiers
     source          1..6598
                     /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    3386..3507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3423..3442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3477..3496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3514..3969)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3651..3672)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3863..3882
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3996..4100
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4102..4459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4102..4293
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4125..4145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4310..4445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4494..5288
                     /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             4494..5288
                     /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(4548..4567)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5158..5177
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5476..5495)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5520..5567
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5896..6484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6385..6404
                     /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 cacattcgat gtgcccactg gctcctccat
     3061 gctgagtatc tacggcccga tttgccccaa gagcagtacg accggcaact ggaacaactt
     3121 tgatctggga tatggcggct tcaagagtga cagaaacgat agcttcaatg ccaataagca
     3181 gcaacagccg atgtggggca ggaaaccgca actgatgtcg gaagacagtt tcgacagtgg
     3241 gatcaaggtg gactagggat ccaccggatc tagataactg atcataatca gccataccac
     3301 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3361 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3421 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3481 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     3541 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3601 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3661 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3721 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3781 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3841 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3901 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3961 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4081 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4141 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4261 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4321 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4381 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4441 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4501 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4561 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4621 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4681 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4741 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4801 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4861 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4921 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4981 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5041 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5101 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5161 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5221 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5281 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5341 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5401 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5461 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5521 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5581 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5641 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5701 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5761 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5821 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6001 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6541 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
