LOCUS       sequence-291089-        6298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin2_501-1191.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6298)
  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 6298)
  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_501-1191_AE
FEATURES             Location/Qualifiers
     source          1..6298
                     /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    3086..3207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3123..3142)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3177..3196
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3214..3669)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3351..3372)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3563..3582
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3696..3800
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3802..4159
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3802..3993
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3825..3845)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4010..4145
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4194..4988
                     /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             4194..4988
                     /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(4248..4267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4858..4877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5176..5195)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5220..5267
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5596..6184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6085..6104
                     /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 aacagtgttt cccagctaat
      901 ctcacgtagt actgacagta ccttaagagc tctggagacc gtgaagaaag tgggaaaggt
      961 tggcgctaat ggtcagaatg ctgctgggcc ctctgcagat tctgtaactg aaaataaaat
     1021 tggttctcca cccaagactc ctgtaagtaa tgtagcagct acctcagctg ggccctctaa
     1081 tgttggaaca gagctgaatt ctgtgcctca aaaatccagc ccatttctaa ctagagtacc
     1141 agtatatcct ccgcattctg aaaacattca gtattttcaa gatccaagga ctcagatacc
     1201 ctttgaagtc ccacagtacc cacagacagg atactatcca ccacctccaa cggtaccagc
     1261 tggtgtggct ccctgtgttc ctcgctttgt gaggtccaat aacgttccag agtcctccct
     1321 cccacctgct tccatgccat atgccgatca ttacagtaca ttttcccctc gagatcgaat
     1381 gaattcttct ccttaccagc ctcctcctcc gcagccgtat ggaccagttc ctccagtacc
     1441 ttctggaatg tatgctcctg tgtacgacag caggcgcatc tggcgcccac ctatgtacca
     1501 acgagatgac attattagaa gcaattcttt acctccaatg gatgtgatgc actcatctgt
     1561 ctatcagaca tctttgcggg aaagatataa ctcattagat ggatattatt cggtggcttg
     1621 tcagccacca agtgagccaa ggacaactgt gcctttacca agggaacctt gtggtcattt
     1681 gaagaccagt tgcgaggagc agataagaag aaagccagat cagtgggcac agtaccacac
     1741 tcagaaagca cctcttgtct cttcaactct tcctgtggca acacagtcac caacaccacc
     1801 ttctcctctg ttcagtgtag actttcgtgc ggatttctca gagagtgtga gtggtacaaa
     1861 atttgaagaa gatcatcttt cccattattc tccctggtct tgtggcacca taggctcctg
     1921 tataaatgcc attgattcag agcccaaaga tgtcattgct aattcaaatg ctgtgttaat
     1981 ggacctggac agtggtgatg ttaagagaag agtacattta tttgaaaccc agagaaggac
     2041 aaaagaagaa gatccaataa ttccctttag tgatggaccc atcatctcaa aatggggtgc
     2101 gatttccaga tcttcccgta caggttacca taccacagat cctgtccagg ccactgcttc
     2161 ccaaggaagt gcgactaagc ccatcagtgt atcagattat gtcccttatg tcaatgctgt
     2221 tgattcaagg tggagttcat atggcaacga ggccacatca tcagcacact atgttgaaag
     2281 ggacagattc attgttactg atttatctgg tcatagaaag cattccagta ctggggacct
     2341 tttgagcctt gaacttcagc aggccaagag caactcatta cttcttcaga gagaggccaa
     2401 tgctttggcc atgcaacaga agtggaattc cctggatgaa ggccgtcacc ttaccttaaa
     2461 ccttttaagc aaggaaattg aactaagaaa tggagagtta cagagtgatt atacagaaga
     2521 tgcaacagat actaaacctg atagggatat cgagttagag ctttcagcac ttgatactga
     2581 tgaacctgat ggacaaagtg aaccaattga agagatcttg gacatacagc ttggtatcag
     2641 ttctcaaaat gatcagttgc taaatggaat ggcagtggaa aatgggcatc cagtacagca
     2701 gcaccaaaag gagccaccaa agcagaagaa acagagttta ggtgaagacc atgtgattct
     2761 ggaggagcaa aaaacaattc tgccggtaac ttcttgcttt agccagccac tcccagtgtc
     2821 tattagcaat gcaagttgcc tccccatcac cacatctgtc agtgctggca acctcattct
     2881 gaaaactcat gttatgtctg aagataaaaa cgacttttta aaacctgttg caaatgggaa
     2941 gatggttaac agctgaggat ccaccggatc tagataactg atcataatca gccataccac
     3001 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3061 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3121 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3181 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     3241 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3301 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3361 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3421 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3481 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3541 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3601 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3661 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3721 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3781 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3841 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3901 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4021 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4081 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4141 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4201 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4261 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4321 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4381 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4441 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4501 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4561 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4621 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4681 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4741 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4801 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4861 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4921 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4981 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5041 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5101 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5161 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5221 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5281 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5341 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5401 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5461 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5521 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5701 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5881 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5941 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6001 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6061 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6121 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6181 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6241 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
