LOCUS       sequence-291081-        6574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmRoquin-del725-755.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6574)
  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 6574)
  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-DmRoquin-del725-755_AE
FEATURES             Location/Qualifiers
     source          1..6574
                     /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    3362..3483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3399..3418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3453..3472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3490..3945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3627..3648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3839..3858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3972..4076
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4078..4435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4078..4269
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4101..4121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4286..4421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4470..5264
                     /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             4470..5264
                     /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(4524..4543)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5134..5153
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5452..5471)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5496..5543
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5872..6460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6361..6380
                     /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 cttatatgcc gattcaggct ccctcatgga cagattttct
      901 gaactgtccc atctgctgca atgaattcgc ggccagccag cgatgtccgg tcagtctggg
      961 ctgcgggcac accatctgca agctatgcct gacgaccctc tacaaccggc agtgtccctt
     1021 cgatcagacc gtgattgtca gtgacatcga caatttgccc atcaaccatg ccttgctcca
     1081 gttggtcaag gactcggagc tcctcgagct ggcaccacca ccgcccagtg tgcagaagct
     1141 ggagccggag caccttaagt gttaccaact gggtcagcgg tgcatcgagg agctggcttt
     1201 gcacttgaag tccttcctca acctgaacgg aaatggcaat cttctaactc gacccatgtt
     1261 acggaaactg gttactctgg tcaactgcca gctgatggag gaggagggac gggtgagagc
     1321 gcttcgagca gccagatccc taggcgaacg cacagtcacc gagctgatcc tgcagcacca
     1381 aaatccccag cagttgagct caaatctgtg ggctgcagtg cggacaagag gatgccagtt
     1441 ccttggaccc gccatgcagg aggaggtgct caaactggtg ctgctggcct tggaggaggg
     1501 atcagcgctt tcgcgcaaag tgctggtcat gttcgtcgtg cagcgactag agccgaagtt
     1561 tccccaggcc tccaagacga gtatcggaca tgtggtacag ctgctctacc gcgccagctg
     1621 cttcaaggtg tccaagcggg aggcggactc gtcgttgatg cagctcaaag aggaattccg
     1681 tacgtatgac gccctacgcc gggagcacga tgcccaaatc gttcagatag ccaccgaggc
     1741 gggtctgcga attgcaccgg aacaatggtc ttccctattg tatggcgacg tgatgcataa
     1801 aagccacatg cagagcatca tcgataagct gcagacgcca agctcctttg cgcaatcagt
     1861 ccaggaattg gtcatcgcct tgcagcggac cagcgatcct gcgaagctat ccagtttgca
     1921 tcatcatctg aagtacctgg ccaatattga tccctgtgcg gaggttgccc cgtggtcggt
     1981 tttggctgag gccttggatg cagtgcggca ttcggtcgtt ggactggtga acttccttca
     2041 gcaccacgga gtgcgcaagg cgcaggacgg catcagtggc ggtggcagtg gtgggacgac
     2101 caatagcaat ccgaagtaca agatcagctt atgcagggac ttgaatgtgc ggcgggtgtg
     2161 tccccgaggt tctagctgta cctttgccca ttcccaggag gaggtagaac gctatcgtgc
     2221 acggaatcga ggaaaacata tgaagacacc gttggcgctg cagggacctc cagcggtagg
     2281 cgtaggagcc ataaagaaac cactaggcga gcaggaggga ccgcctctag gtaacatgcc
     2341 acccatgtta cctatgtcgc ctatgcatta tatgggctct cctaggggat atctggatcc
     2401 cagcttggga ctttcgccag gtggtggtct cccgccgtcg catcacagtc ccattaccag
     2461 gctcatagtg ccgagccgct acgattcgcg ctttagcggg tttggcggtg gcacgcccag
     2521 gattccttcg cccagagaat accaagccaa tccggtagcg ccaacgcaac gcaatgccaa
     2581 cccaccgaat tttagtgtga acagtaacct gcacaagggc tacatgttgc ccgccagcgg
     2641 tggagatgtc ttccatctgg ccaatccgtg ggagcaagca tacctggcac agcagcaaca
     2701 tcctccacag catccccaac agcagcagcc accgtccagc aagccgaatc ccagccgacc
     2761 gttgtcgatt ttacccgcaa cagctgatac ctcatttttt gaaaagaaac cgccgaatag
     2821 tgttagcata gatctggaca gggtcccgga ggtcaatgtg gatgcagtgc ctctgttccg
     2881 gcgatccaac aataaccaca ataacaacaa tagtaactcc cacaataaca acaacaacaa
     2941 tcatggctca tcactgctgt tctggaacaa tacgggtaaa gattcagcca actttgtacg
     3001 atcagactcc atattggatg acgatgcgtc cacattctat ggcggcttca agagtgacag
     3061 aaacgatagc ttcaatgcca ataagcagca acagccgatg tggggcagga aaccgcaact
     3121 gatgtcggaa gacagtttcg agggcggcat cgatagtgga atgatgctgc agctggagaa
     3181 gaatctggtg gacatcgtcg atcccgatga cagtgggatc aaggtggact agggatccac
     3241 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3301 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3361 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3421 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3481 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3541 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3601 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3661 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3721 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3781 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3841 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3901 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3961 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4021 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4081 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4141 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4201 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4261 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4321 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4381 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4441 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4501 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4561 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4621 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4681 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4741 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4801 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4861 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4921 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4981 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5041 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5101 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5161 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5221 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5281 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5341 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5401 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5461 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5521 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5581 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5641 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5701 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5761 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5821 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5881 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5941 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6001 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6061 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6121 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6181 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6241 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6301 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6361 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6421 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6481 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6541 tgattctgtg gataaccgta ttaccgccat gcat
//
