LOCUS       sequence-290070-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of DmNot2-del344-380.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  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 5667)
  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-del344-380_T
FEATURES             Location/Qualifiers
     source          1..5667
                     /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    2455..2576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2492..2511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2546..2565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2583..3038)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2720..2741)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2932..2951
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3065..3169
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3171..3528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3171..3362
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3194..3214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3379..3514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3563..4357
                     /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             3563..4357
                     /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(3617..3636)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4227..4246
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4545..4564)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4589..4636
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4965..5553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5454..5473
                     /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 tgacttaaca acattaggcc tcaatctgaa
     1861 ctctcctgaa aatctctacc ccaaatttgc gtcaccctgg gcatcttcac cttgtcgacc
     1921 tcaagacata gacttccatg ttccatctga gtacttaacg aacattcaca ttagggataa
     1981 gctggctgca ataaaacttg gccgatatgg tgaagacctt ctcttctatc tctattacat
     2041 gaatggagga gacgtattac aacttttagc tgcagtggag ctttttaacc gtgattggag
     2101 ataccacaaa gaagaacgag tatggattac cagggcacca ggcatggagc caacaatgaa
     2161 aaccaatacc tatgagaggg gaacatatta cttctttgac tgtcttaact ggaggaaagt
     2221 agctaaggag ttccatctgg aatatgacaa attagaagaa cggcctcacc tgccatccac
     2281 cttcaactac aaccctgctc agcaagcctt ctaaccgcgg gcccgggatc caccggatct
     2341 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2401 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2461 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2521 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     2581 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2641 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2701 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2761 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2821 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2881 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2941 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3001 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3061 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3121 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3181 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3541 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3601 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3661 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3721 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3781 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3841 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3901 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3961 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4021 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4081 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4141 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4201 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4261 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4321 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4381 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4441 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4501 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4561 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4621 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4681 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4741 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4801 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4861 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4921 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4981 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5041 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5101 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5161 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5221 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5281 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5341 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5401 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5461 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5521 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5581 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5641 gtggataacc gtattaccgc catgcat
//
