LOCUS       sequence-291090-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin1_1-500.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5727)
  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 5727)
  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-HsRoquin_1-500_AE
FEATURES             Location/Qualifiers
     source          1..5727
                     /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    2515..2636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2552..2571)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2606..2625
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2643..3098)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2780..2801)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2992..3011
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3125..3229
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3231..3588
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3231..3422
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3254..3274)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3439..3574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3623..4417
                     /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             3623..4417
                     /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(3677..3696)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4287..4306
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4605..4624)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4649..4696
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5025..5613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5514..5533
                     /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 aatgcctgta caagctccac aatggacgga
      901 tttcctctcc tgcccaattt gcactcagac tttcgacgaa acaattcgaa agcccatcag
      961 tttgggttgt ggccatactg tctgcaagat gtgcctgaat aaactccacc gcaaggcttg
     1021 cccatttgac cagaccacta tcaatacaga cattgagctc ctccctgtga actcagcatt
     1081 gctgcagctc gtgggtgctc aggtccctga gcagcagcct attactttgt gtagtggggt
     1141 tgaagacaca aagcattatg aggaagccaa gaaatgtgta gaagaattag cattgtactt
     1201 aaaaccgctc agcagtgcta gaggagtggg tctgaacagc actactcaga gtgttctgag
     1261 tcgcccaatg cagaggaagc ttgtgactct agtccactgt caactagtag aagaagaagg
     1321 caggattcgt gccatgaggg cagctcgatc tttaggtgaa cgaacagtta cagagctcat
     1381 tctccagcac cagaatcctc agcaactctc ttccaatctt tgggcagcag taagggctag
     1441 gggatgccag ttccttggac cagcaatgca ggaggaagct ttaaagttgg ttctgctggc
     1501 cttagaagat ggttctgctt tgtcaagaaa agtattggtt ctgtttgtgg tgcaaagatt
     1561 ggagccacgg tttcctcaag cctctaaaac tagcattggg catgttgttc agctccttta
     1621 tagagcctcc tgtttcaagg tcaccaaacg tgatgaagac tcttctttga tgcagctgaa
     1681 agaagaattt agaacctatg aagctctgcg gcgagaacat gactcccaga tagtgcagat
     1741 tgctatggaa gcaggcttac gaattgcacc ggaccagtgg tcctctttgc tttatggaga
     1801 ccagtctcac aaatctcata tgcagtccat tattgacaag ttgcagactc cagcctcttt
     1861 tgcacagagt gttcaggaac taacaattgc tctccagcga actggagacc cagcaaactt
     1921 gaaccgacta agaccccatt tggagctgtt agcaaacatt gaccctagtc cagatgctcc
     1981 tcctcctact tgggaacagc tggaaaatgg gctggttgct gtgcgtacag tggtacatgg
     2041 gctggttgat tatatccaga accacagcaa aaaaggagca gatcagcagc agcctccaca
     2101 gcatagcaaa tacaaaacat acatgtgtcg agatatgaag cagagaggag gatgccctcg
     2161 tggggccagc tgtacatttg cacactcaca ggaggaactg gaaaaatttc gtaaaatgaa
     2221 taagcgcctg gttccgagaa gacccttgag tgcctctttg ggtcaactta atgaggtggg
     2281 cctgccttca gcagctatcc ttccagatga aggtgcagtg gatctcccta gcagaaaacc
     2341 tcctgctctg ccaaatggaa ttgtatcaac ataaccgcgg gcccgggatc caccggatct
     2401 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2461 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2521 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2581 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     2641 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2701 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2761 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2821 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2881 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2941 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3001 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3061 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3121 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3181 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3241 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3301 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3421 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3481 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3541 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3601 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3661 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3721 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3781 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3841 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3901 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3961 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4021 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4081 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4141 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4201 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4261 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4321 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4381 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4441 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4501 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4561 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4621 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4681 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4741 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4801 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4861 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4921 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4981 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5041 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5101 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5161 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5221 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5281 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5341 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5401 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5461 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5521 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5581 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5641 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5701 gtggataacc gtattaccgc catgcat
//
