LOCUS       sequence-291087-        7798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  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 7798)
  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_AE
FEATURES             Location/Qualifiers
     source          1..7798
                     /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    4586..4707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4623..4642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4677..4696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4714..5169)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4851..4872)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5063..5082
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5196..5300
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5302..5659
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5302..5493
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5325..5345)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5510..5645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5694..6488
                     /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             5694..6488
                     /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(5748..5767)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6358..6377
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6676..6695)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6720..6767
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7096..7684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7585..7604
                     /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 aacagtgttt cccagctaat
     2401 ctcacgtagt actgacagta ccttaagagc tctggagacc gtgaagaaag tgggaaaggt
     2461 tggcgctaat ggtcagaatg ctgctgggcc ctctgcagat tctgtaactg aaaataaaat
     2521 tggttctcca cccaagactc ctgtaagtaa tgtagcagct acctcagctg ggccctctaa
     2581 tgttggaaca gagctgaatt ctgtgcctca aaaatccagc ccatttctaa ctagagtacc
     2641 agtatatcct ccgcattctg aaaacattca gtattttcaa gatccaagga ctcagatacc
     2701 ctttgaagtc ccacagtacc cacagacagg atactatcca ccacctccaa cggtaccagc
     2761 tggtgtggct ccctgtgttc ctcgctttgt gaggtccaat aacgttccag agtcctccct
     2821 cccacctgct tccatgccat atgccgatca ttacagtaca ttttcccctc gagatcgaat
     2881 gaattcttct ccttaccagc ctcctcctcc gcagccgtat ggaccagttc ctccagtacc
     2941 ttctggaatg tatgctcctg tgtacgacag caggcgcatc tggcgcccac ctatgtacca
     3001 acgagatgac attattagaa gcaattcttt acctccaatg gatgtgatgc actcatctgt
     3061 ctatcagaca tctttgcggg aaagatataa ctcattagat ggatattatt cggtggcttg
     3121 tcagccacca agtgagccaa ggacaactgt gcctttacca agggaacctt gtggtcattt
     3181 gaagaccagt tgcgaggagc agataagaag aaagccagat cagtgggcac agtaccacac
     3241 tcagaaagca cctcttgtct cttcaactct tcctgtggca acacagtcac caacaccacc
     3301 ttctcctctg ttcagtgtag actttcgtgc ggatttctca gagagtgtga gtggtacaaa
     3361 atttgaagaa gatcatcttt cccattattc tccctggtct tgtggcacca taggctcctg
     3421 tataaatgcc attgattcag agcccaaaga tgtcattgct aattcaaatg ctgtgttaat
     3481 ggacctggac agtggtgatg ttaagagaag agtacattta tttgaaaccc agagaaggac
     3541 aaaagaagaa gatccaataa ttccctttag tgatggaccc atcatctcaa aatggggtgc
     3601 gatttccaga tcttcccgta caggttacca taccacagat cctgtccagg ccactgcttc
     3661 ccaaggaagt gcgactaagc ccatcagtgt atcagattat gtcccttatg tcaatgctgt
     3721 tgattcaagg tggagttcat atggcaacga ggccacatca tcagcacact atgttgaaag
     3781 ggacagattc attgttactg atttatctgg tcatagaaag cattccagta ctggggacct
     3841 tttgagcctt gaacttcagc aggccaagag caactcatta cttcttcaga gagaggccaa
     3901 tgctttggcc atgcaacaga agtggaattc cctggatgaa ggccgtcacc ttaccttaaa
     3961 ccttttaagc aaggaaattg aactaagaaa tggagagtta cagagtgatt atacagaaga
     4021 tgcaacagat actaaacctg atagggatat cgagttagag ctttcagcac ttgatactga
     4081 tgaacctgat ggacaaagtg aaccaattga agagatcttg gacatacagc ttggtatcag
     4141 ttctcaaaat gatcagttgc taaatggaat ggcagtggaa aatgggcatc cagtacagca
     4201 gcaccaaaag gagccaccaa agcagaagaa acagagttta ggtgaagacc atgtgattct
     4261 ggaggagcaa aaaacaattc tgccggtaac ttcttgcttt agccagccac tcccagtgtc
     4321 tattagcaat gcaagttgcc tccccatcac cacatctgtc agtgctggca acctcattct
     4381 gaaaactcat gttatgtctg aagataaaaa cgacttttta aaacctgttg caaatgggaa
     4441 gatggttaac agctgaggat ccaccggatc tagataactg atcataatca gccataccac
     4501 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4561 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4621 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4681 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     4741 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4801 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4861 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4921 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4981 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5041 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5101 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5161 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5221 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5281 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5341 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5401 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5461 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5521 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5581 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5641 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5701 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5761 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5821 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5881 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5941 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6001 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6061 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6121 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6181 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     6241 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6301 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6361 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6421 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6481 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6541 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6601 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6661 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6721 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6781 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6841 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6901 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6961 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7021 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7081 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7201 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     7261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7681 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7741 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
