LOCUS       sequence-291088-        5725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin2_1-500.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5725)
  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 5725)
  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-HsRoquin2_1-500_AE
FEATURES             Location/Qualifiers
     source          1..5725
                     /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    2513..2634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2550..2569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2604..2623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2641..3096)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2778..2799)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2990..3009
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3123..3227
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3229..3586
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3229..3420
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3252..3272)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3437..3572
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3621..4415
                     /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             3621..4415
                     /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(3675..3694)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4285..4304
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4603..4622)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4647..4694
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5023..5611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5512..5531
                     /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 cttcgaattc tgcagtcgac atgcctgtgc aggcagctca
      901 atggacagaa tttctgtcct gtccaatctg ctataatgaa tttgatgaga atgtgcacaa
      961 acccatcagt ttaggttgtt cacacactgt ttgcaagacc tgcttgaata aacttcatcg
     1021 aaaagcttgt ccttttgacc agactgccat caacacagat attgatgtac ttcctgtcaa
     1081 cttcgcactt ctccagttag ttggagccca ggtaccagat catcagtcaa ttaagttaag
     1141 taatctaggt gagaataaac actatgaggt tgcaaagaaa tgcgttgagg atttggcact
     1201 ctacttaaaa ccactaagtg gaggtaaagg tgtagctagc ttgaaccaga gtgcactgag
     1261 ccgtccaatg caaaggaaac tggtgacact tgtaaactgt caactggtgg aggaagaagg
     1321 tcgtgtaaga gccatgcgag cagctcgttc ccttggagaa agaactgtaa cagaactgat
     1381 attacagcac cagaaccctc agcagttgtc tgccaatcta tgggccgctg tcagggctcg
     1441 aggatgccag tttttagggc cagctatgca agaagaggcc ttgaagctgg tgttactggc
     1501 attagaagat ggttctgccc tctcaaggaa agttctggta ctttttgttg tgcagagact
     1561 agaaccaaga tttcctcagg catcaaaaac aagtattggt catgttgtgc aactactgta
     1621 tcgagcttct tgttttaagg ttaccaaaag agatgaagac tcttccctaa tgcagctgaa
     1681 ggaggaattt cggagttatg aagcattacg cagagaacat gatgcccaaa ttgttcatat
     1741 tgccatggaa gcaggactcc gtatttcacc tgaacagtgg tcctctcttt tgtatggtga
     1801 tttggctcat aaatcacaca tgcagtctat cattgataag ctacagtctc cagagtcatt
     1861 tgcaaagagt gtccaggaat tgacaattgt tttgcaacga acaggtgacc cagctaactt
     1921 aaatagactg aggcctcatt tagagcttct tgcaaacata gaccctaatc cagacgctgt
     1981 ttcaccaact tgggagcagc tggaaaatgc aatggtagct gttaaaacag tagttcatgg
     2041 ccttgtggac ttcatacaaa attatagtag aaaaggccat gagacccctc agcctcagcc
     2101 aaacagcaaa tacaagacta gcatgtgccg agatttgcga cagcaagggg gttgtccacg
     2161 aggaacaaat tgtacatttg cccattctca ggaagagctt gaaaagtatc gattaaggaa
     2221 caaaaagatc aatgccactg taagaacgtt tcctcttcta aataaagttg gtgtaaacaa
     2281 cactgtcaca accacagccg gaaatgtcat ttctgtcata ggaagtactg aaacaacagg
     2341 gaaaattgtt ccaagtacaa acggaatttc aaatgcagaa tgaggatcca ccggatctag
     2401 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2461 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2521 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2581 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2641 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2701 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2761 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2821 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2881 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2941 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3001 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3061 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3121 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3181 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3241 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3301 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3421 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3481 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3541 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3601 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3661 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3721 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3781 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3841 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3901 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3961 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4021 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4081 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4141 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4201 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4261 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4321 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4381 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4441 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4501 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4561 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4621 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4681 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4741 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4801 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4861 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4921 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4981 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5041 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5101 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5161 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5221 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5281 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5341 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5401 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5461 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5521 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5581 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5641 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5701 ggataaccgt attaccgcca tgcat
//
