LOCUS       sequence-360167-        5050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5050)
  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 5050)
  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-HsNot8_J
FEATURES             Location/Qualifiers
     source          1..5050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             605..670
                     /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             695..721
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1815..1834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1906..2361)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2043..2064)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2255..2274
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2388..2492
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2494..2851
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2494..2685
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2517..2537)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2702..2837
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2886..3680
                     /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             2886..3680
                     /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(2940..2959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3550..3569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3868..3887)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3912..3959
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4288..4876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4777..4796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      661 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      721 tcctccatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gccctgcagc
      781 acttgtggag aatagccagg ttatctgtga agtgtgggcc agtaatctag aagaagagat
      841 gaggaagatc cgagaaatcg tgctcagtta cagttatatt gccatggaca cagaatttcc
      901 aggtgttgtg gtgcgaccaa ttggtgaatt tcgtagttcc atagattacc aatatcagct
      961 tctgcggtgc aatgttgacc ttttaaaaat tatccagctg ggccttacat tcacaaatga
     1021 gaagggagag tatccttctg gaatcaatac ttggcagttc aatttcaaat ttaaccttac
     1081 agaggacatg tactcccagg attccataga tctccttgct aactcaggac tacagtttca
     1141 gaagcatgaa gaggaaggga ttgacacact gcactttgca gagctgctta tgacatcagg
     1201 agtggttctc tgtgacaatg tcaaatggct ttcatttcat agtggctatg attttggcta
     1261 tatggtaaag ttgcttacag attctcgttt gccagaagag gaacatgaat tctttcatat
     1321 tctgaacctt ttcttcccat ccatttatga tgtgaaatac ctgatgaaga gctgcaaaaa
     1381 tcttaaggga ggtcttcagg aagttgctga tcagttggat ttgcagagga ttggaaggca
     1441 gcaccaggca ggctcagact cactgctgac aggaatggct ttctttagga tgaaagagtt
     1501 gttttttgag gacagcattg atgatgccaa gtactgtggg cggctctatg gcttaggcac
     1561 aggagtggcc cagaagcaga atgaggatgt ggactctgcc caggagaaga tgagcatcct
     1621 ggcgattatc aacaacatgc agcagtgagg atccaccgga tctagataac tgatcataat
     1681 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     1921 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1981 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2041 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2101 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2161 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2221 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2281 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2341 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2401 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2461 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2521 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2581 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2641 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2701 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2761 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2821 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2881 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2941 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3001 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3061 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3121 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3181 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3241 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3301 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3361 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3421 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3481 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3541 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3601 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3661 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3721 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3781 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3841 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3901 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3961 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4021 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4081 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4141 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4201 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4261 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4321 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4501 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4621 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4681 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4861 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4921 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4981 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     5041 tggagttccg
//
