LOCUS       sequence-291085-        7626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  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 7626)
  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-HsRoquin1_AE
FEATURES             Location/Qualifiers
     source          1..7626
                     /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"
     RBS             3076..3084
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4414..4535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4451..4470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4505..4524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4542..4997)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4679..4700)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4891..4910
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5024..5128
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5130..5487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5130..5321
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5153..5173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5338..5473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5522..6316
                     /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             5522..6316
                     /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(5576..5595)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6186..6205
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6504..6523)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6548..6595
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6924..7512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7413..7432
                     /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 agggaataca gtaacacagc ttattccgcg
     2401 agggacagac cccagctatg attctagtct gaaaccagga aaaatagatc atctgagcag
     2461 tagtgctcct ggatcccctc ctgacctgct agaatctgtt cctaagagta tttctgcctt
     2521 acctgtgaat ccacattcta tacctccaag ggggccagca gatctgcctc caatgcctgt
     2581 taccaaacca cttcagatgg tacctcgagg ttctcagtta tatccagcac aacagacgga
     2641 tgtttattat caggatcctc gaggagcagc tccgccattt gaaccagcac cttatcagca
     2701 gggtatgtat tatactccac caccacaatg tgtgtcccgc tttgtccgac ctccaccatc
     2761 tgctcctgaa cctgctcctc cctacttgga tcattatcca ccctacctcc aagaacgtgt
     2821 tgtaaactct cagtatggca cacagccaca gcagtaccca cctatatacc catctcacta
     2881 tgatggccgt cgagtgtacc ctgctccgtc ttacacaaga gaagagatat tccgagaaag
     2941 ccctataccc attgagattc cacctgcagc agtaccatcg tatgtaccag aatccagaga
     3001 aagataccaa cagatcgaga gttactatcc agtggctcct catccaactc agatcagacc
     3061 ttcgtacctc agagaacctc cttatagccg gcttcctcct cctccccagc ctcatcctag
     3121 tctagatgaa ctacatcgcc gacgaaagga aataatggcc cagctagagg aaagaaaggt
     3181 tatctctcca cctccttttg caccttcacc aaccttgcct cctacctttc atccggaaga
     3241 atttttggat gaagacttga aggtagctgg gaaatacaaa ggaaatgatt atagccaata
     3301 ctctccctgg tcatgtgaca ccatcggctc ctacattgga accaaagatg caaaacccaa
     3361 agatgttgtg gcagcaggga gtgtggaaat gatgaatgtg gagagtaaag gaatgaggga
     3421 ccagcgatta gatcttcaga gaagagcagc agaaaccagt gatgatgacc tcatcccatt
     3481 tggagaccga ccaacagtgt ctcggtttgg tgccatctca cgaacttcca aaactatata
     3541 tcagggtgct ggtccaatgc aggctatggc acctcaggga gctcctacaa aatctattaa
     3601 catttcagat tatagtccat atggaaccca cggtggctgg ggagcttctc catattcacc
     3661 tcatcaaaac ataccttctc agggacactt cagtgagagg gagagaatat ctatgtcaga
     3721 agtggccagt catggaaaac cccttccatc tgctgagaga gaacagctac gactagaatt
     3781 gcagcaattg aaccatcaga ttagccagca gacccagcta cgtggactag aggctgttag
     3841 taacaggctg gtgttgcaga gggaggcaaa caccctggca ggccagtcac agcccccacc
     3901 accgccacct ccaaaatggc ctgggatgat ctcaagtgag cagttgagct tggaactgca
     3961 ccaggtggaa agggaaatcg ggaagagaac acgggaactg agtatggaaa accagtgttc
     4021 tctggacatg aaaagcaaac tgaatacaag taaacaagca gaaaatggac aaccagaacc
     4081 acaaaacaag gttccggctg aggaccttac attgacattc agtgatgtac caaatggatc
     4141 agccttgaca caagagaata tcagcctcct atcaaacaag accagctctc tgaacctgtc
     4201 agaggaccct gagggaggag gggataataa tgactcccag agatcaggag ttactcccag
     4261 ttctgctcct taaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     4321 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4381 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4441 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4501 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     4561 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4621 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4681 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4741 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4801 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4861 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4921 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4981 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5041 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5101 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     5161 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5221 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5281 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5401 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5461 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5521 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5581 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5641 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5701 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5761 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5821 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5881 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5941 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     6001 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     6061 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     6121 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6181 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6241 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6301 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6361 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6421 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6481 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6541 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6601 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6661 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6721 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6781 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6841 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     7081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7621 atgcat
//
