LOCUS       sequence-420971-        7745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin1_510-1133.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7745)
  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 7745)
  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-HsRoquin1_510-1133_AE
FEATURES             Location/Qualifiers
     source          1..7745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        344..547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     498..518
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     592..611
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        592..610
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1062..1088
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     RBS             1820..1828
                     /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)"
     promoter        complement(3027..3045)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3028..3045)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3065..3082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3071..3295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3341..3769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3428..3447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3638..3659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3778..3798)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3783..4112
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3963..4098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4025..4044
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4179..4973
                     /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(4233..4252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4843..4862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5147..5268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5184..5203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5238..5257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5317..5333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5317..5333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5330..5352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5341..5357
                     /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(5365..5371,5372..5389,5390..5395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5410..5431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5548..5565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5719..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5799..5818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..7269,7270..7338))
                     /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     7101..7120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7339..7443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7518..7537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7709..7745,1..343)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
       61 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      301 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      361 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      421 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      481 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      541 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
      601 tcactatagg gagacccaag cttgccacca tggcttctaa ctttactcag ttcgttctcg
      661 tcgacaatgg cggaactggc gacgtgactg tcgccccaag caacttcgct aacggggtcg
      721 ctgaatggat cagctctaac tcgcgatcac aggcttacaa agtaacctgt agcgttcgtc
      781 agagctctgc gcagaatcgc aaatacacca tcaaagtcga ggtgcctaaa gtggcaaccc
      841 agactgttgg tggtgaagag cttcctgtag ccggatggag atcttactta aatatggaac
      901 taaccattcc aattttcgcc acgaattccg actgcgagct tattgttaag gcaatgcaag
      961 gtctcctaaa agatggaaac ccgattccct cggccatcgc ggccaactcc ggcatctacg
     1021 gaggtggagg tggatctggt ggatctcctc cacatatggc ttacccatac gatgttccag
     1081 attacgctcc tccaaagctt ggatccgaat tcccgggtcg actcgagtct agagcggccg
     1141 cccgagggac agaccccagc tatgattcta gtctgaaacc aggaaaaata gatcatctga
     1201 gcagtagtgc tcctggatcc cctcctgacc tgctagaatc tgttcctaag agtatttctg
     1261 ccttacctgt gaatccacat tctatacctc caagggggcc agcagatctg cctccaatgc
     1321 ctgttaccaa accacttcag atggtacctc gaggttctca gttatatcca gcacaacaga
     1381 cggatgttta ttatcaggat cctcgaggag cagctccgcc atttgaacca gcaccttatc
     1441 agcagggtat gtattatact ccaccaccac aatgtgtgtc ccgctttgtc cgacctccac
     1501 catctgctcc tgaacctgct cctccctact tggatcatta tccaccctac ctccaagaac
     1561 gtgttgtaaa ctctcagtat ggcacacagc cacagcagta cccacctata tacccatctc
     1621 actatgatgg ccgtcgagtg taccctgctc cgtcttacac aagagaagag atattccgag
     1681 aaagccctat acccattgag attccacctg cagcagtacc atcgtatgta ccagaatcca
     1741 gagaaagata ccaacagatc gagagttact atccagtggc tcctcatcca actcagatca
     1801 gaccttcgta cctcagagaa cctccttata gccggcttcc tcctcctccc cagcctcatc
     1861 ctagtctaga tgaactacat cgccgacgaa aggaaataat ggcccagcta gaggaaagaa
     1921 aggttatctc tccacctcct tttgcacctt caccaacctt gcctcctacc tttcatccgg
     1981 aagaattttt ggatgaagac ttgaaggtag ctgggaaata caaaggaaat gattatagcc
     2041 aatactctcc ctggtcatgt gacaccatcg gctcctacat tggaaccaaa gatgcaaaac
     2101 ccaaagatgt tgtggcagca gggagtgtgg aaatgatgaa tgtggagagt aaaggaatga
     2161 gggaccagcg attagatctt cagagaagag cagcagaaac cagtgatgat gacctcatcc
     2221 catttggaga ccgaccaaca gtgtctcggt ttggtgccat ctcacgaact tccaaaacta
     2281 tatatcaggg tgctggtcca atgcaggcta tggcacctca gggagctcct acaaaatcta
     2341 ttaacatttc agattatagt ccatatggaa cccacggtgg ctggggagct tctccatatt
     2401 cacctcatca aaacatacct tctcagggac acttcagtga gagggagaga atatctatgt
     2461 cagaagtggc cagtcatgga aaaccccttc catctgctga gagagaacag ctacgactag
     2521 aattgcagca attgaaccat cagattagcc agcagaccca gctacgtgga ctagaggctg
     2581 ttagtaacag gctggtgttg cagagggagg caaacaccct ggcaggccag tcacagcccc
     2641 caccaccgcc acctccaaaa tggcctggga tgatctcaag tgagcagttg agcttggaac
     2701 tgcaccaggt ggaaagggaa atcgggaaga gaacacggga actgagtatg gaaaaccagt
     2761 gttctctgga catgaaaagc aaactgaata caagtaaaca agcagaaaat ggacaaccag
     2821 aaccacaaaa caaggttccg gctgaggacc ttacattgac attcagtgat gtaccaaatg
     2881 gatcagcctt gacacaagag aatatcagcc tcctatcaaa caagaccagc tctctgaacc
     2941 tgtcagagga ccctgaggga ggaggggata ataatgactc ccagagatca ggagttactc
     3001 ccagttctgc tccttaaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     3061 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3121 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3181 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3241 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3301 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3361 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3421 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3481 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3541 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3601 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3661 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3721 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3781 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3961 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4021 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4081 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4141 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4201 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4261 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4321 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4381 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4441 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4501 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4561 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4621 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4681 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4741 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4801 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4981 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5041 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5101 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5161 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5221 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5281 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7501 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7561 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7621 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     7681 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     7741 tcaat
//
