LOCUS       sequence-291064-        7920 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 7920)
  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 7920)
  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: pcDNA3.1-MS2-HA-HsRoquin2_501-1191_AE
FEATURES             Location/Qualifiers
     source          1..7920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3476..3494)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3477..3494)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3514..3531)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3520..3744
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3790..4218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3877..3896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4087..4108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4227..4247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4232..4561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4412..4547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4474..4493
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4628..5422
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4682..4701)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5292..5311
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5596..5717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5633..5652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5687..5706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5766..5782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5766..5782)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5779..5801)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5790..5806
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5814..5820,5821..5838,5839..5844))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5859..5880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5997..6014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6168..6753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6248..6267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6924..7715,7716..7784))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7547..7566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7785..7889)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccaac
     1381 agtgtttccc agctaatctc acgtagtact gacagtacct taagagctct ggagaccgtg
     1441 aagaaagtgg gaaaggttgg cgctaatggt cagaatgctg ctgggccctc tgcagattct
     1501 gtaactgaaa ataaaattgg ttctccaccc aagactcctg taagtaatgt agcagctacc
     1561 tcagctgggc cctctaatgt tggaacagag ctgaattctg tgcctcaaaa atccagccca
     1621 tttctaacta gagtaccagt atatcctccg cattctgaaa acattcagta ttttcaagat
     1681 ccaaggactc agataccctt tgaagtccca cagtacccac agacaggata ctatccacca
     1741 cctccaacgg taccagctgg tgtggctccc tgtgttcctc gctttgtgag gtccaataac
     1801 gttccagagt cctccctccc acctgcttcc atgccatatg ccgatcatta cagtacattt
     1861 tcccctcgag atcgaatgaa ttcttctcct taccagcctc ctcctccgca gccgtatgga
     1921 ccagttcctc cagtaccttc tggaatgtat gctcctgtgt acgacagcag gcgcatctgg
     1981 cgcccaccta tgtaccaacg agatgacatt attagaagca attctttacc tccaatggat
     2041 gtgatgcact catctgtcta tcagacatct ttgcgggaaa gatataactc attagatgga
     2101 tattattcgg tggcttgtca gccaccaagt gagccaagga caactgtgcc tttaccaagg
     2161 gaaccttgtg gtcatttgaa gaccagttgc gaggagcaga taagaagaaa gccagatcag
     2221 tgggcacagt accacactca gaaagcacct cttgtctctt caactcttcc tgtggcaaca
     2281 cagtcaccaa caccaccttc tcctctgttc agtgtagact ttcgtgcgga tttctcagag
     2341 agtgtgagtg gtacaaaatt tgaagaagat catctttccc attattctcc ctggtcttgt
     2401 ggcaccatag gctcctgtat aaatgccatt gattcagagc ccaaagatgt cattgctaat
     2461 tcaaatgctg tgttaatgga cctggacagt ggtgatgtta agagaagagt acatttattt
     2521 gaaacccaga gaaggacaaa agaagaagat ccaataattc cctttagtga tggacccatc
     2581 atctcaaaat ggggtgcgat ttccagatct tcccgtacag gttaccatac cacagatcct
     2641 gtccaggcca ctgcttccca aggaagtgcg actaagccca tcagtgtatc agattatgtc
     2701 ccttatgtca atgctgttga ttcaaggtgg agttcatatg gcaacgaggc cacatcatca
     2761 gcacactatg ttgaaaggga cagattcatt gttactgatt tatctggtca tagaaagcat
     2821 tccagtactg gggacctttt gagccttgaa cttcagcagg ccaagagcaa ctcattactt
     2881 cttcagagag aggccaatgc tttggccatg caacagaagt ggaattccct ggatgaaggc
     2941 cgtcacctta ccttaaacct tttaagcaag gaaattgaac taagaaatgg agagttacag
     3001 agtgattata cagaagatgc aacagatact aaacctgata gggatatcga gttagagctt
     3061 tcagcacttg atactgatga acctgatgga caaagtgaac caattgaaga gatcttggac
     3121 atacagcttg gtatcagttc tcaaaatgat cagttgctaa atggaatggc agtggaaaat
     3181 gggcatccag tacagcagca ccaaaaggag ccaccaaagc agaagaaaca gagtttaggt
     3241 gaagaccatg tgattctgga ggagcaaaaa acaattctgc cggtaacttc ttgctttagc
     3301 cagccactcc cagtgtctat tagcaatgca agttgcctcc ccatcaccac atctgtcagt
     3361 gctggcaacc tcattctgaa aactcatgtt atgtctgaag ataaaaacga ctttttaaaa
     3421 cctgttgcaa atgggaagat ggttaacagc tgagcggccg cctagagggc cctattctat
     3481 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3541 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3601 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3661 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3721 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3781 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4201 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4261 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4321 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4381 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4441 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4501 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4561 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4621 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4681 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4741 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4801 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4861 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4921 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4981 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5041 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5101 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5161 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5221 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5281 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5341 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5401 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5461 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5521 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5581 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5641 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5701 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5761 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5821 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5881 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5941 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6001 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6061 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     6721 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6781 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6841 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6901 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6961 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7021 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7081 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7141 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7201 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7261 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7321 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7381 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7441 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7501 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7561 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7621 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7681 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7741 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7801 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7861 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
