LOCUS       sequence-291060-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRoquin1_1-509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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_1-509_AE
FEATURES             Location/Qualifiers
     source          1..7374
                     /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(2930..2948)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2931..2948)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2968..2985)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2974..3198
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3244..3672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3331..3350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3541..3562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3681..3701)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3686..4015
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3866..4001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3928..3947
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4082..4876
                     /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(4136..4155)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4746..4765
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5050..5171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5087..5106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5141..5160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5220..5236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5220..5236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5233..5255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5244..5260
                     /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(5268..5274,5275..5292,5293..5298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5313..5334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5451..5468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5622..6207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5702..5721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6378..7169,7170..7238))
                     /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     7001..7020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7239..7343)
                     /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 tggatccatg
     1381 cctgtacaag ctccacaatg gacggatttc ctctcctgcc caatttgcac tcagactttc
     1441 gacgaaacaa ttcgaaagcc catcagtttg ggttgtggcc atactgtctg caagatgtgc
     1501 ctgaataaac tccaccgcaa ggcttgccca tttgaccaga ccactatcaa tacagacatt
     1561 gagctcctcc ctgtgaactc agcattgctg cagctcgtgg gtgctcaggt ccctgagcag
     1621 cagcctatta ctttgtgtag tggggttgaa gacacaaagc attatgagga agccaagaaa
     1681 tgtgtagaag aattagcatt gtacttaaaa ccgctcagca gtgctagagg agtgggtctg
     1741 aacagcacta ctcagagtgt tctgagtcgc ccaatgcaga ggaagcttgt gactctagtc
     1801 cactgtcaac tagtagaaga agaaggcagg attcgtgcca tgagggcagc tcgatcttta
     1861 ggtgaacgaa cagttacaga gctcattctc cagcaccaga atcctcagca actctcttcc
     1921 aatctttggg cagcagtaag ggctagggga tgccagttcc ttggaccagc aatgcaggag
     1981 gaagctttaa agttggttct gctggcctta gaagatggtt ctgctttgtc aagaaaagta
     2041 ttggttctgt ttgtggtgca aagattggag ccacggtttc ctcaagcctc taaaactagc
     2101 attgggcatg ttgttcagct cctttataga gcctcctgtt tcaaggtcac caaacgtgat
     2161 gaagactctt ctttgatgca gctgaaagaa gaatttagaa cctatgaagc tctgcggcga
     2221 gaacatgact cccagatagt gcagattgct atggaagcag gcttacgaat tgcaccggac
     2281 cagtggtcct ctttgcttta tggagaccag tctcacaaat ctcatatgca gtccattatt
     2341 gacaagttgc agactccagc ctcttttgca cagagtgttc aggaactaac aattgctctc
     2401 cagcgaactg gagacccagc aaacttgaac cgactaagac cccatttgga gctgttagca
     2461 aacattgacc ctagtccaga tgctcctcct cctacttggg aacagctgga aaatgggctg
     2521 gttgctgtgc gtacagtggt acatgggctg gttgattata tccagaacca cagcaaaaaa
     2581 ggagcagatc agcagcagcc tccacagcat agcaaataca aaacatacat gtgtcgagat
     2641 atgaagcaga gaggaggatg ccctcgtggg gccagctgta catttgcaca ctcacaggag
     2701 gaactggaaa aatttcgtaa aatgaataag cgcctggttc cgagaagacc cttgagtgcc
     2761 tctttgggtc aacttaatga ggtgggcctg ccttcagcag ctatccttcc agatgaaggt
     2821 gcagtggatc tccctagcag aaaacctcct gctctgccaa atggaattgt atcaacaggg
     2881 aatacagtaa cacagcttat tccgtgagcg gccgcctaga gggccctatt ctatagtgtc
     2941 acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3001 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3061 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3121 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3181 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3241 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3301 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3361 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3421 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3481 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3541 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3601 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3661 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3781 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3841 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3901 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3961 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4021 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4081 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4141 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4201 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4261 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4321 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4381 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4441 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4501 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4561 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4621 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4681 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4741 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4801 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4861 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4921 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4981 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5041 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5101 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5161 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5281 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     6181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6661 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6961 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
