LOCUS       sequence-289251-        7401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1-1089.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  AUTHORS   Petit AP, Wohlbold L, Bawankar P, Huntzinger E, Schmidt S, 
            Izaurralde E, Weichenrieder O
  TITLE     The structural basis for the interaction between the CAF1 nuclease 
            and the NOT1 scaffold of the human CCR4-NOT deadenylase complex.
  JOURNAL   Nucleic Acids Res. 2012 Nov;40(21):11058-72. doi: 
            10.1093/nar/gks883. Epub 2012 Sep 12.
  PUBMED    22977175
REFERENCE   2  (bases 1 to 7401)
  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: 
            "10.1093/nar/gks883"; journalName: "Nucleic Acids Res"; date: 
            "2012-11"; volume: "40"; issue: "21"; pages: "11058-72"
COMMENT     Sequence Label: pLNHA-C1-HsNot1iso1_1-1089_K
FEATURES             Location/Qualifiers
     source          1..7401
                     /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    4189..4310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4226..4245)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4280..4299
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4317..4772)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4454..4475)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4666..4685
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4799..4903
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4905..5262
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4905..5096
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4928..4948)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5113..5248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5297..6091
                     /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             5297..6091
                     /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(5351..5370)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5961..5980
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6279..6298)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6323..6370
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6699..7287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7188..7207
                     /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 cgagctaatc ttgactcgct ctcgctggcc ttgtctcaaa tcagctacct
      841 ggtggacaat ttaaccaaga aaaattaccg agccagccag caggaaatac agcatattgt
      901 gaatcggcac ggtcctgagg cagacaggca tttattacgc tgcctatttt cgcatgtgga
      961 tttcagtggc gatggtaaaa gcagtggcaa agatttccat cagactcagt ttctgattca
     1021 ggagtgtgcg ttgctgatta caaagccaaa ttttatctcg acgctgtcct atgccattga
     1081 taatccattg cactatcaga agagtttaaa gcctgcaccc cacttatttg cccagctgag
     1141 taaagtgctc aaattaagca aagtacaaga ggtaattttt ggccttgccc tgttgaattc
     1201 ttccagctca gatcttagag gtttcgctgc ccagtttatc aaacagaagc ttccagatct
     1261 tctgcgttct tacattgacg cagacgtcag tggaaatcaa gaaggtggct tccaagatat
     1321 agcaatagag gtcctacacc tcctcctctc ccatctcctc tttgggcaga agggagcctt
     1381 tggagttgga caagaacaga tagacgcttt tcttaagacg ctgcgcagag attttcccca
     1441 agaacgctgt cccgtggtgc tcgcaccact tttataccct gaaaaacggg acattctaat
     1501 ggacaggatc ctgcctgatt ccggaggggt agctaaaacc atgatggaga gctctttggc
     1561 tgatttcatg caagaagtag gctatggctt ttgtgcaagt attgaagaat gtcgcaatat
     1621 aatcgtgcag tttggtgttc gggaggtcac agctgcccag gttgcaaggg ttttgggaat
     1681 gatggctcga actcattcag gattaacaga tggcattcca ttacagagta tttctgctcc
     1741 gggcagtggg atctggagtg atgggaaaga taaaagtgat ggagcacagg cacacacatg
     1801 gaatgtagaa gtcttgattg acgttcttaa agaactgaat ccaagtttga atttcaagga
     1861 agtaacttat gaactggacc atcctggatt tcaaattcgt gacagtaaag gacttcataa
     1921 tgtggtttat ggcattcaga ggggtttggg tatggaagtg ttcccagtag acctcatata
     1981 tagaccttgg aaacatgctg aaggccagct ctccttcatt caacattccc ttataaatcc
     2041 agagatcttc tgttttgctg actatccctg tcatactgtt gccactgata ttctgaaagc
     2101 accaccagag gatgacaatc gagaaattgc cacatggaag agcttggatt tgattgaatc
     2161 tctgctgagg cttgcagagg ttgggcagta tgagcaagtc aaacagctct tcagcttccc
     2221 tatcaaacac tgtccagaca tgctggtatt ggccttacta caaattaaca cctcttggca
     2281 taccttgcgc catgaactta tctccactct gatgccaatt ttccttggaa accatcctaa
     2341 ctcagctatt attttgcact atgcatggca tgggcaggga cagtctccct caattcgcca
     2401 acttatcatg catgcaatgg cagaatggta catgagaggg gagcagtatg atcaggccaa
     2461 attgtctcga atacttgatg tggcccagga cttgaaggcc ttgtcaatgc tgctaaatgg
     2521 tactccattt gcctttgtta ttgaccttgc tgcacttgct tcacgtcgtg aatacctcaa
     2581 acttgataag tggctcacag ataaaattcg agagcatggg gagcctttta tccaggcgtg
     2641 tatgactttt ttaaagagac ggtgtccttc tattttgggc ggacttgccc cagaaaaaga
     2701 ccagcccaaa agtgctcaac ttcctccaga aactttggcg acaatgttgg cctgtctgca
     2761 agcttgtgca gggagtgttt ctcaggagct atcagaaact atcctcacca tggtagccaa
     2821 ttgcagtaat gttatgaata aggccagaca accaccacct ggagttatgc caaaaggacg
     2881 tcctcctagt gctagcagct tagatgccat ttctcctgtt cagattgacc ctcttgctgg
     2941 aatgacatct cttagtatag gtggttcagc tgcccctcac acccagagta tgcagggttt
     3001 tcctccaaat ttgggttctg cattcagtac ccctcagtca ccagcaaaag catttccacc
     3061 cctttcaacc cccaatcaga ccactgcatt cagtggtatt ggaggacttt catcacagct
     3121 tccagtaggt ggtcttggca caggcagcct gactggtata ggaactggtg ctcttggact
     3181 ccctgcagtg aataacgacc cttttgtaca gaggaaactg ggcacctctg gactgaatca
     3241 gcctacattc cagcagacgg acttgtctca ggtgtggcca gaggcaaacc agcactttag
     3301 taaagagata gatgatgaag caaacagcta tttccagcga atatataatc atccaccaca
     3361 tccaaccatg tctgttgatg aggtattaga aatgctgcag agatttaaag actctactat
     3421 aaagagggaa cgagaagtat ttaactgtat gctaaggaac ttgtttgaag aatatcgttt
     3481 ttttccccag tatcctgata aagagttaca tataacagcc tgcctatttg gtggtataat
     3541 tgagaaagga ctggtcactt acatggcact aggtctggct ctacgatatg ttcttgaagc
     3601 cttacgcaag ccttttggat ccaaaatgta ttatttcggg attgctgcac tagatagatt
     3661 taaaaacaga ttgaaggact atccccagta ttgtcagcat ttggcttcta tcagtcactt
     3721 tatgcaattt ccacatcatt tacaggagta tattgagtat ggacagcagt ctagagatcc
     3781 tcctgtgaaa atgcaaggct ctatcacaac ccctggaagt attgcactgg ctcaggccca
     3841 ggctcaggcc caggttccag caaaagctcc tcttgctggt caagttagca ctatggtaac
     3901 cacctcaaca actaccactg ttgctaaaac ggttacggtc accaggccaa ctggagtcag
     3961 ctttaagaaa gatgtgccac cttctattaa tactacaaat atagatacgt tgcttgtggc
     4021 cacagatcaa actgagagaa ttgtgtgacc gcgggcccgg gatccaccgg atctagataa
     4081 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4141 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4201 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4261 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     4321 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4381 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4441 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4501 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4561 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4621 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4681 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4741 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4801 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4861 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4921 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4981 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5041 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5101 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5161 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5221 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     5281 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5341 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5401 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5461 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5521 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5581 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5641 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5701 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5761 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5821 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5881 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5941 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6001 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6061 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6121 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6181 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6241 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     6301 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     6361 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6421 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6481 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6541 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6601 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6661 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6721 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6781 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6841 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6901 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6961 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7021 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7081 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7141 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7201 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7261 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7321 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7381 aaccgtatta ccgccatgca t
//
