LOCUS       sequence-289218-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  AUTHORS   Braun JE, Huntzinger E, Fauser M, Izaurralde E
  TITLE     GW182 proteins directly recruit cytoplasmic deadenylase complexes to
            miRNA targets.
  JOURNAL   Mol Cell. 2011 Oct 7;44(1):120-33.
  PUBMED    21981923
REFERENCE   2  (bases 1 to 6370)
  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: "Mol Cell.";
            date: "2011-10-7"; volume: "44(1)"; pages: "120-33"
COMMENT     Sequence Label: pLNHA-C1-HsNot10_K
FEATURES             Location/Qualifiers
     source          1..6370
                     /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             665..730
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3158..3279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3195..3214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3249..3268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3286..3741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3423..3444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3635..3654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3768..3872
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3874..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3874..4065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4266..5060
                     /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             4266..5060
                     /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(4320..4339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4930..4949
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5248..5267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5292..5339
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5668..6256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6157..6176
                     /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 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctgctg cagacaagcc tgcagatcag ggagcagaga aacatgaagg
      841 cacaggtcag tcctctggga tcactgatca agagaaggag ttatccacca atgctttcca
      901 agctttcaca tctggaaatt atgatgcctg tctacaacac cttgcctgtc tacaagatat
      961 aaacaaagat gattataaaa taattttgaa tacagcagta gctgagtttt ttaaaagtaa
     1021 ccaaacaaca acagataatt tgagacaaac acttaaccag ctgaagaatc aggtccactc
     1081 agctgttgaa gaaatggatg gattagatga tgttgaaaac agcatgttgt actataatca
     1141 agcagtcatt ctttatcatc tgcggcagta tacagaagcc atatcagttg gtgaaaaact
     1201 ttatcagttc atagagcctt ttgaagaaaa atttgcccaa gcagtgtgtt ttttgcttgt
     1261 agacctgtat atattaacct accaagctga gaaagctttg catcttcttg ctgtcctaga
     1321 aaaaatgatt tcacagggta acaataacaa aaatggaaag aatgagactg gtaataacaa
     1381 caacaaagat ggatctaatc ataaagctga aagtggagct ctaatagaag ctgcaaaatc
     1441 aaagatacat cagtacaaag tacgagctta tatccaaatg aagtctctga aagcatgtaa
     1501 aagggaaatc aagtcagtca tgaatacagc tggaaattcc gcaccctctc tctttcttaa
     1561 aagcaatttt gagtacttaa gaggtaatta tcgaaaagcc gtgaagctat taaatagttc
     1621 aaacattgct gagcatccag gattcatgaa aacaggtgaa tgcttgagat gcatgttctg
     1681 gaataacctt ggttgcatcc attttgccat gagcaagcac aatttgggaa tattctactt
     1741 taaaaaggct ctgcaagaga atgacaatgt ctgtgcacag ctcagtgcag gtagcactga
     1801 tccaggtaaa aaattttcag gaagacccat gtgtacgtta ctaaccaata agagatatga
     1861 gttgctgtat aactgtggaa ttcagcttct tcacattgga aggcctcttg ctgccttcga
     1921 atgtctgatt gaagctgttc aggtttatca tgcaaatcct cgcctctggc tacggctggc
     1981 tgaatgctgc attgctgcca ataaggggac ttctgaacaa gaaactaaag gccttcccag
     2041 caaaaaagga attgtacagt ctattgttgg tcaaggctat catcgtaaaa tagttttggc
     2101 atcacagtct atacagaata ctgtttataa tgatggtcag tcttcggcca ttcctgtagc
     2161 cagtatggag tttgcagcca tatgtctcag aaatgccttg ttgctgctac ctgaagaaca
     2221 gcaagatcca aagcaggaaa atggggctaa aaatagtaat caattaggtg ggaacacaga
     2281 gagcagcgaa agcagtgaaa cttgcagcag taaaagccat gatggagata aattcattcc
     2341 agctccacct tcttctccat tgagaaaaca ggaattagaa aacttaaagt gctccatact
     2401 tgcttgcagt gcctacgtgg ctctggcttt gggtgataac ctcatggctt tgaatcatgc
     2461 agataaactt cttcagcagc ccaagctgtc aggatctctt aagtttttgg gacatttata
     2521 tgctgcagaa gccctcatct ctctcgacag aatatctgat gccattactc acttgaaccc
     2581 ggagaatgtc actgatgtct ccttagggat ctcttcaaat gagcaggacc aaggatcaga
     2641 caaaggtgaa aatgaagcaa tggaatcctc tggtaagcgg gcccctcagt gctaccccag
     2701 ttccgtcaac tctgccagga ctgtgatgct gttcaacctt ggcagcgctt actgcctgag
     2761 gagcgaatat gacaaagccc gaaagtgtct ccaccaggcg gcttcaatga tccatcctaa
     2821 agaggtgccc cctgaggcca tcttgctggc agtctacctt gaactgcaga atggtaatac
     2881 tcagctggcc ttacagatca tcaaaaggaa tcagctgctc cctgcagtga aaacacactc
     2941 tgaagtgaga aagaagccag tgtttcagcc tgtccacccg atccagccca tccaaatgcc
     3001 ggctttcacc actgtgcaga gaaagtgagg atccaccgga tctagataac tgatcataat
     3061 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3121 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     3301 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3361 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3421 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3481 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3541 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3601 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3661 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3721 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3781 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3841 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4261 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4321 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4381 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4441 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4501 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4561 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4621 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4681 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4741 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4801 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4861 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4921 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4981 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5041 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5101 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5161 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5221 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5281 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5341 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5401 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5461 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6301 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6361 cgccatgcat
//
