LOCUS       sequence-289957-        5778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot2-W507E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5778)
  AUTHORS   Boland A, Chen Y, Raisch T, Jonas S, Kuzuoglu-Ozturk D, Wohlbold L, 
            Weichenrieder O, Izaurralde E
  TITLE     Structure and assembly of the NOT module of the human CCR4-NOT 
            complex.
  JOURNAL   Nat Struct Mol Biol. 2013 Nov;20(11):1289-97. doi: 
            10.1038/nsmb.2681. Epub 2013 Oct 13.
  PUBMED    24121232
REFERENCE   2  (bases 1 to 5778)
  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"; doi: "10.1038/nsmb.2681";
            journalName: "Nat Struct Mol Biol"; date: "2013-11"; volume: "20"; 
            issue: "11"; pages: "1289-97"
COMMENT     Sequence Label: pLNHA-C1-HsNot2-W507E_S
FEATURES             Location/Qualifiers
     source          1..5778
                     /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    2566..2687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2603..2622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2657..2676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2694..3149)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2831..2852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3043..3062
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3176..3280
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3282..3639
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3282..3473
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3305..3325)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3490..3625
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3674..4468
                     /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             3674..4468
                     /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(3728..3747)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4338..4357
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4656..4675)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4700..4747
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5076..5664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5565..5584
                     /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 cgagctcaag cttcggtgag gactgatgga catacattat ctgagaaaag
      841 aaactaccag gtgacaaaca gcatgtttgg tgcttcaaga aagaagtttg tagagggggt
      901 cgacagtgac taccatgacg aaaacatgta ctacagccag tcttctatgt ttccacatcg
      961 gtcagaaaaa gatatgctgg catcaccatc tacatcaggt cagctgtctc agtttggggc
     1021 aagtttatac gggcaacaaa gtgcactagg ccttccaatg agggggatga gcaacaatac
     1081 ccctcagtta aatcgcagct tatcacaagg cactcagtta ccgagccacg tcacgccaac
     1141 aacaggggta ccaacaatgt cacttcacac gcctccatct ccaagcaggg gtattttgcc
     1201 tatgaatcct aggaatatga tgaaccactc ccaggttggt cagggcattg gaattcctag
     1261 caggacaaat agcatgagca gttcagggtt aggtagcccc aacagaagct cgccaagcat
     1321 aatatgtatg ccaaagcagc agccttctcg acagcctttt actgtgaaca gtatgtctgg
     1381 atttggaatg aacaggaatc aggcatttgg aatgaataac tccttatcaa gtaacatttt
     1441 taatggaaca gacggaagtg aaaatgtgac aggattggac ctttcagatt tcccagcatt
     1501 agcagaccga aacaggaggg aaggaagtgg taacccaact ccattaataa accccttggc
     1561 tggaagagct ccttatgttg gaatggtaac aaaaccagca aatgaacaat cccaggactt
     1621 ctcaatacac aatgaagatt ttccagcatt accaggctcc agctataaag atccaacatc
     1681 aagtaatgat gacagtaaat ctaatttgaa tacatctggc aagacaactt caagtacaga
     1741 tggacccaaa ttccctggag ataaaagttc aacaacacaa aataataacc agcagaaaaa
     1801 agggatccag gtgttacctg atggtcgggt tactaacatt cctcaaggga tggtgacgga
     1861 ccaatttgga atgattggcc tgttaacatt tatcagggca gcagagacag acccaggaat
     1921 ggtacatctt gcattaggaa gtgacttaac aacattaggc ctcaatctga actctcctga
     1981 aaatctctac cccaaatttg cgtcaccctg ggcatcttca ccttgtcgac ctcaagacat
     2041 agacttccat gttccatctg agtacttaac gaacattcac attagggata agctggctgc
     2101 aataaaactt ggccgatatg gtgaagacct tctcttctat ctctattaca tgaatggagg
     2161 agacgtatta caacttttag ctgcagtgga gctttttaac cgtgattgga gataccacaa
     2221 agaagaacga gtatggatta ccagggcacc aggcatggag ccaacaatga aaaccaatac
     2281 ctatgagagg ggaacatatt acttctttga ctgtcttaac gagaggaaag tagctaagga
     2341 gttccatctg gaatatgaca aattagaaga acggcctcac ctgccatcca ccttcaacta
     2401 caaccctgct cagcaagcct tctaaccgcg ggcccgggat ccaccggatc tagataactg
     2461 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2521 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2581 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2641 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     2701 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2761 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2821 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2881 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2941 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3001 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3061 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3121 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3181 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3241 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3301 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3361 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3481 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3541 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3601 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3661 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3721 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3781 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3841 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3901 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3961 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4021 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4081 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4141 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4201 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4261 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4321 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4381 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4441 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4501 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4561 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4621 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4681 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4741 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4801 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4861 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5221 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5761 cgtattaccg ccatgcat
//
