LOCUS       sequence-290446-        6423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of 
            HsNot1iso1_840-1605-del1097-1110-IKGRQtoDSYAA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  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 6423)
  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: 
            pLNHA-C1-HsNot1iso1_840-1605-del1097-1110-IKGRQtoDSYAA_X
FEATURES             Location/Qualifiers
     source          1..6423
                     /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    3211..3332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3248..3267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3302..3321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3339..3794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3476..3497)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3688..3707
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3821..3925
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3927..4284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3927..4118
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3950..3970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4135..4270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4319..5113
                     /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             4319..5113
                     /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(4373..4392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4983..5002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5301..5320)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5345..5392
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5721..6309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6210..6229
                     /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 cgagccatgt ttagtaaaga gatagatgat gaagcaaaca gctatttcca
      841 gcgaatatat aatcatccac cacatccaac catgtctgtt gatgaggtat tagaaatgct
      901 gcagagattt aaagactcta ctataaagag ggaacgagaa gtatttaact gtatgctaag
      961 gaacttgttt gaagaatatc gtttttttcc ccagtatcct gataaagagt tacatataac
     1021 agcctgccta tttggtggta taattgagaa aggactggtc acttacatgg cactaggtct
     1081 ggctctacga tatgttcttg aagccttacg caagcctttt ggatccaaaa tgtattattt
     1141 cgggattgct gcactagata gatttaaaaa cagattgaag gactatcccc agtattgtca
     1201 gcatttggct tctatcagtc actttatgca atttccacat catttacagg agtatattga
     1261 gtatggacag cagtctagag atcctcctgt gaaaatgcaa ggctctatca caacccctgg
     1321 aagtattgca ctggctcagg cccaggctca ggcccaggtt ccagcaaaag ctcctcttgc
     1381 tggtcaagtt agcactatgg taaccacctc aacaactacc actgttgcta aaacggttac
     1441 ggtcaccagg ccaactggag tcagctttaa gaaagatgtg ccaccttcta ttaatactac
     1501 aaatatagat acgttgcttg tggccacaga tcaaactgag agaattgtgg agcccccaga
     1561 aaatatccag ggtagtagtg gtatgacaca aaaggttgaa gagctaaagg aaacggtgaa
     1621 agaagaattt atgccttggg tttcacagta tctggttatg aagagagtca gtattgagcc
     1681 aaactttcat agcctgtatt caaacttcct tgacacgctg aagaatcctg aatttaacaa
     1741 gatggttctg aatgagacct acagaaacat taaagtgctc ctgacctctg ataaagctgc
     1801 agccaatttc tcagatcgtt ctttgctgaa gaacttggga cattggctag gaatgatcac
     1861 attagctaaa aacaaaccca tcttacacac tgacttggat gtgaaatcat tgctgctaga
     1921 ggcttatgtt aaaggacaac aagaattgct ctatgtagtg ccctttgttg ccaaagtctt
     1981 agaatctagc attcgtagtg tggtttttag gccaccaaac ccttggacaa tggcaattat
     2041 gaatgtatta gctgagctac atcaggagca tgacttaaag ttaaacttga agtttgaaat
     2101 cgaggttctc tgcaagaacc ttgcattaga catcaatgag ctaaaacctg gaaacctcct
     2161 aaaggataaa gatcgcctga agaatttaga tgagcaactc tctgctccaa agaaagatgt
     2221 caagcagcca gaagaactcc ctcccatcac aaccacaaca acttctacta caccagctac
     2281 caacaccact tgtacagcca cggttccacc acagccacag tacagctacc acgacatcaa
     2341 tgtctattcc cttgcgggct tggcaccaca cattactcta aatccaacaa ttcccttgtt
     2401 tcaggcccat ccacagttga agcagtgtgt gcgtcaggca attgaacggg ctgtccagga
     2461 gctggtccat cctgtggtgg atcgatcaat taagattgcc atgactactt gtgagcaaga
     2521 tgtcaggtcg gattttgccc tggattcgga ggaatctcga atgcgaatag cagctcatca
     2581 catgatgcgt aacttgacag cttacatggc tatgattaca tgcgctgaac ctttgctcat
     2641 gagcatatct accaacttaa aaaacagttt tgcctcagcc cttcgtactg cttccccaca
     2701 acaaagagaa atgatggatc aggcagctgc tcaattagct caggacaatt gtgagttggc
     2761 ttgctgtttt attcagaaga ctgcagtaga aaaagcaggc cctgagatgg acaagagatt
     2821 agcaactgaa tttgagctga gaaaacatgc taggcaagaa ggacgcagat actgtgatcc
     2881 tgttgtttta acatatgctg ctgaacggat gccagagcaa atcaggctga aagttggtgg
     2941 tgtggaccca aagcagttgg ctgtttacga agagtttgca cgcaatgttc ctggcttctt
     3001 acctacaaat gacttaagtc agcccacggg atttttagcc cagcccatga agcaagcttg
     3061 ggcaacatag ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     3121 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3181 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3241 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3301 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3361 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3421 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3481 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3541 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3601 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3661 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3721 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3781 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3841 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3901 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3961 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4081 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4141 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4201 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4261 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4501 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5101 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5161 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5221 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5281 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5341 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5401 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5461 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5521 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5581 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5641 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5701 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5761 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5821 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5881 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5941 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6001 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6061 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6121 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6181 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6241 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6301 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6361 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6421 cat
//
