LOCUS       sequence-291084-        6667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin-ILLVtoEEEE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  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 6667)
  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-ILLVtoEEEE_AE
FEATURES             Location/Qualifiers
     source          1..6667
                     /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    3455..3576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3492..3511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3546..3565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3583..4038)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3720..3741)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3932..3951
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4065..4169
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4171..4528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4171..4362
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4194..4214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4379..4514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4563..5357
                     /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             4563..5357
                     /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(4617..4636)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5227..5246
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5545..5564)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5589..5636
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5965..6553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6454..6473
                     /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 tcgagggcgg
     3241 cgaagatagt ggaatgatgc tgcaggaaga gaagaatgaa gtggacatcg aagatcccga
     3301 tgacagtggg atcaaggtgg actagggatc caccggatct agataactga tcataatcag
     3361 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3421 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3481 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3541 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3601 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3661 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3721 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3781 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3841 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3901 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3961 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4021 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4141 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4201 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4321 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4381 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4441 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4501 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4561 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4621 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4681 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4741 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4801 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4861 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4921 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4981 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5041 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5101 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5161 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5221 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5281 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5341 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5401 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5461 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5521 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5581 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5641 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5701 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5761 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5821 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5881 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5941 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6001 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6061 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6121 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6181 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6241 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6301 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6361 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6421 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6481 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6541 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6601 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6661 catgcat
//
