LOCUS       sequence-290326-        7516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1318-1605.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7516)
  AUTHORS   Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT, 
            Weichenrieder O, Izaurralde E
  TITLE     A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct 
            links between miRNA target recognition and silencing.
  JOURNAL   Mol Cell. 2014 Jun 5;54(5):737-50. doi: 
            10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.
  PUBMED    24768540
REFERENCE   2  (bases 1 to 7516)
  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: 
            "10.1016/j.molcel.2014.03.034"; journalName: "Mol Cell"; date: 
            "2014-06-5- 5"; volume: "54"; issue: "5"; pages: "737-50"
COMMENT     Sequence Label: pCIneo-lambdaN-HsNot1_1318-1605-HA-MBP_W
FEATURES             Location/Qualifiers
     source          1..7516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..1160
                     /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             2052..2078
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3036..3045
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3220..3341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3231..3250)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3285..3304
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3527..3982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3614..3633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3824..3845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4099..4456
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4099..4290
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4122..4142)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4307..4442
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4507..5301
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4561..4580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5171..5190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5365..5413
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5454..5473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5573..5595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5633..5651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5719..5823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5824..5892,5893..6684)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6042..6061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6855..7443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7344..7363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag
     1141 ctcaatggaa agctgcaaac ccaccgctcg aggctccaaa gaaagatgtc aagcagccag
     1201 aagaactccc tcccatcaca accacaacaa cttctactac accagctacc aacaccactt
     1261 gtacagccac ggttccacca cagccacagt acagctacca cgacatcaat gtctattccc
     1321 ttgcgggctt ggcaccacac attactctaa atccaacaat tcccttgttt caggcccatc
     1381 cacagttgaa gcagtgtgtg cgtcaggcaa ttgaacgggc tgtccaggag ctggtccatc
     1441 ctgtggtgga tcgatcaatt aagattgcca tgactacttg tgagcaaata gtcaggaagg
     1501 attttgccct ggattcggag gaatctcgaa tgcgaatagc agctcatcac atgatgcgta
     1561 acttgacagc tggaatggct atgattacat gcagggaacc tttgctcatg agcatatcta
     1621 ccaacttaaa aaacagtttt gcctcagccc ttcgtactgc ttccccacaa caaagagaaa
     1681 tgatggatca ggcagctgct caattagctc aggacaattg tgagttggct tgctgtttta
     1741 ttcagaagac tgcagtagaa aaagcaggcc ctgagatgga caagagatta gcaactgaat
     1801 ttgagctgag aaaacatgct aggcaagaag gacgcagata ctgtgatcct gttgttttaa
     1861 catatcaagc tgaacggatg ccagagcaaa tcaggctgaa agttggtggt gtggacccaa
     1921 agcagttggc tgtttacgaa gagtttgcac gcaatgttcc tggcttctta cctacaaatg
     1981 acttaagtca gcccacggga tttttagccc agcccatgaa gcaagcttgg gcaacagcga
     2041 attcaatggc ttacccatac gatgttccag attacgctat gaaaatcgaa gaaggtaaac
     2101 tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc ggtaagaaat
     2161 tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg gaagagaaat
     2221 tcccgcaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca cacgaccgct
     2281 ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg
     2341 acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg attgcttacc
     2401 cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg aacccgccaa
     2461 aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt aagagcgcgc
     2521 tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct gacgggggtt
     2581 atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg gataacgctg
     2641 gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aagcaaacac atgaatgcag
     2701 acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg atgaccatca
     2761 acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt gtaacggtac
     2821 tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta
     2881 acgccgccag tccgaacaaa gagctggcga aagagttcct cgaaaactat ctgctgactg
     2941 atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg ctgaagtctt
     3001 acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac gcccagaaag
     3061 gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg
     3121 tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagat taggcggccg
     3181 cttcccttta gtgagggtta atgcttcgag cagacatgat aagatacatt gatgagtttg
     3241 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3301 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3361 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     3421 acaaatgtgg taaaatccga taaggatcga tccgggctgg cgtaatagcg aagaggcccg
     3481 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     3541 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3601 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3661 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3721 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3781 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3841 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3901 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3961 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4021 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     4081 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4141 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4201 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4321 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4381 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4441 cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taaggctaga
     4501 gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4561 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4621 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4681 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4741 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4801 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4861 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4921 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4981 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5041 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5101 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5161 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5221 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5281 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5341 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     5401 ggttttttgt gtgaatcgat agcgataagg atccgcgtat ggtgcactct cagtacaatc
     5461 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5521 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5581 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5641 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5821 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5881 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5941 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6001 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6061 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6121 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6181 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6241 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6301 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6361 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6421 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6481 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6541 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6601 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6661 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6961 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7501 atggctcgac agatct
//
