LOCUS       sequence-289144-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot6L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  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 5834)
  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-HsNot6L_J
FEATURES             Location/Qualifiers
     source          1..5834
                     /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    2622..2743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2659..2678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2713..2732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2750..3205)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2887..2908)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3099..3118
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3232..3336
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3338..3695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3338..3529
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3361..3381)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3546..3681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3730..4524
                     /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             3730..4524
                     /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(3784..3803)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4394..4413
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4712..4731)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4756..4803
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5132..5720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5621..5640
                     /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 cgagctagac taatagggat gccaaaggaa aaatatgatc ctccagatcc
      841 tcgcagaatt tataccatca tgtcagcaga ggaggtagcc aatgggaaaa aatctcactg
      901 ggcagaatta gaaatctcgg gtagagtgcg gagcctaagt acatcacttt ggtcattgac
      961 acacttgaca gcgctgcacc taaatgacaa ttaccttagt cgcattccac ctgatattgc
     1021 caagcttcat aatctggttt acctggatct gtcatccaat aaactcagaa gtttaccagc
     1081 agaactagga aacatggtgt ctctcaggga attgctttta aataacaatc tgttacgggt
     1141 tttgccttat gaacttggtc ggctcttcca gctacaaact ctaggtttga aaggcaatcc
     1201 tttatcacag gatattctca acttatacca ggacccagat ggaacccgaa agctactgaa
     1261 cttcatgctt gacaatctcg cagttcatcc agagcagctt cctccgaggc catggattac
     1321 attaaaagaa cgagaccaaa ttctgccgtc agcatcattc acggttatgt gttacaatgt
     1381 gttatgtgat aaatacgcta cccggcagct atatggctat tgcccatcct gggcattaaa
     1441 ctgggaatac aggaaaaagg gaattatgga agaaattgtt aactgtgacg cagatatcat
     1501 tagtcttcag gaagtggaaa cagagcaata cttcactctc tttctgccag cattgaagga
     1561 gcgtggatat gatggatttt tttctccaaa gtcacgtgcc aaaatcatgt ctgagcagga
     1621 gagaaagcat gtagatggtt gtgcaatatt cttcaaaaca gaaaaattta cattggtgca
     1681 gaagcataca gtggaattta accaagtggc gatggctaat tcagatggat ccgaagctat
     1741 gctgaacaga gtgatgacaa aagataacat tggtgtcgct gtggtattag aggtccacaa
     1801 agaactattt ggagcaggta tgaagcctat tcatgctgca gacaaacagc tgcttatagt
     1861 ggcaaatgcc cacatgcatt gggacccaga gtattctgat gtgaagctca tccagaccat
     1921 gatgtttgtc tcagaggtta aaaacattct ggagaaagcc tctagtaggc ctggaagccc
     1981 aactgcagat cctaattcca tcccgctggt gctatgtgca gatcttaact cattgccaga
     2041 ttcaggtgtt gtggaatact taagcaatgg aggagtagct gacaaccata aagacttcaa
     2101 ggaactaagg tacaatgagt gtcttatgaa cttcagctgc aatggaaaga atggaagctc
     2161 agaagggaga atcacacatg gcttccaact taagagcgcc tatgaaaata acttgatgcc
     2221 ttacaccaat tacacctttg atttcaaagg cgtgattgac tacattttct attccaagac
     2281 tcatatgaac gtgcttggtg tcctggggcc tttagatcct caatggctgg ttgagaacaa
     2341 catcactggg tgtccacacc ctcacatccc ttcagaccac ttctcactgt taacacaact
     2401 tgaactccac cctccactcc tgcctcttgt caatggtgtt cacttgccta atcggaggta
     2461 ggaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca
     2521 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2581 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2641 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2701 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     2761 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2821 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2881 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2941 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3001 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3061 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3121 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3181 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3241 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3301 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3361 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3601 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3661 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3721 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3781 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3841 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3901 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3961 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4021 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4081 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4141 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4201 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4261 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4321 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4381 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4441 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4501 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4561 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4621 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4681 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4741 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4801 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4861 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4921 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4981 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5041 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5101 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5161 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5221 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5281 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5341 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5401 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5461 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5521 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5581 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5641 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5701 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5761 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5821 ttaccgccat gcat
//
