LOCUS       sequence-393060-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_840-1605.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  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 6453)
  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_X
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             605..670
                     /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             695..721
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3181..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3272..3291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3309..3764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3446..3467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3658..3677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3791..3895
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3897..4254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3897..4088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3920..3940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4105..4240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4289..5083
                     /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             4289..5083
                     /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(4343..4362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4953..4972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5271..5290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5315..5362
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5691..6279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6180..6199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      661 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      721 tcctccatct cgagccatgt ttagtaaaga gatagatgat gaagcaaaca gctatttcca
      781 gcgaatatat aatcatccac cacatccaac catgtctgtt gatgaggtat tagaaatgct
      841 gcagagattt aaagactcta ctataaagag ggaacgagaa gtatttaact gtatgctaag
      901 gaacttgttt gaagaatatc gtttttttcc ccagtatcct gataaagagt tacatataac
      961 agcctgccta tttggtggta taattgagaa aggactggtc acttacatgg cactaggtct
     1021 ggctctacga tatgttcttg aagccttacg caagcctttt ggatccaaaa tgtattattt
     1081 cgggattgct gcactagata gatttaaaaa cagattgaag gactatcccc agtattgtca
     1141 gcatttggct tctatcagtc actttatgca atttccacat catttacagg agtatattga
     1201 gtatggacag cagtctagag atcctcctgt gaaaatgcaa ggctctatca caacccctgg
     1261 aagtattgca ctggctcagg cccaggctca ggcccaggtt ccagcaaaag ctcctcttgc
     1321 tggtcaagtt agcactatgg taaccacctc aacaactacc actgttgcta aaacggttac
     1381 ggtcaccagg ccaactggag tcagctttaa gaaagatgtg ccaccttcta ttaatactac
     1441 aaatatagat acgttgcttg tggccacaga tcaaactgag agaattgtgg agcccccaga
     1501 aaatatccag gagaaaattg cttttatttt caataatctc tcacagtcaa atatgacaca
     1561 aaaggttgaa gagctaaagg aaacggtgaa agaagaattt atgccttggg tttcacagta
     1621 tctggttatg aagagagtca gtattgagcc aaactttcat agcctgtatt caaacttcct
     1681 tgacacgctg aagaatcctg aatttaacaa gatggttctg aatgagacct acagaaacat
     1741 taaagtgctc ctgacctctg ataaagctgc agccaatttc tcagatcgtt ctttgctgaa
     1801 gaacttggga cattggctag gaatgatcac attagctaaa aacaaaccca tcttacacac
     1861 tgacttggat gtgaaatcat tgctgctaga ggcttatgtt aaaggacaac aagaattgct
     1921 ctatgtagtg ccctttgttg ccaaagtctt agaatctagc attcgtagtg tggtttttag
     1981 gccaccaaac ccttggacaa tggcaattat gaatgtatta gctgagctac atcaggagca
     2041 tgacttaaag ttaaacttga agtttgaaat cgaggttctc tgcaagaacc ttgcattaga
     2101 catcaatgag ctaaaacctg gaaacctcct aaaggataaa gatcgcctga agaatttaga
     2161 tgagcaactc tctgctccaa agaaagatgt caagcagcca gaagaactcc ctcccatcac
     2221 aaccacaaca acttctacta caccagctac caacaccact tgtacagcca cggttccacc
     2281 acagccacag tacagctacc acgacatcaa tgtctattcc cttgcgggct tggcaccaca
     2341 cattactcta aatccaacaa ttcccttgtt tcaggcccat ccacagttga agcagtgtgt
     2401 gcgtcaggca attgaacggg ctgtccagga gctggtccat cctgtggtgg atcgatcaat
     2461 taagattgcc atgactactt gtgagcaaat agtcaggaag gattttgccc tggattcgga
     2521 ggaatctcga atgcgaatag cagctcatca catgatgcgt aacttgacag ctggaatggc
     2581 tatgattaca tgcagggaac ctttgctcat gagcatatct accaacttaa aaaacagttt
     2641 tgcctcagcc cttcgtactg cttccccaca acaaagagaa atgatggatc aggcagctgc
     2701 tcaattagct caggacaatt gtgagttggc ttgctgtttt attcagaaga ctgcagtaga
     2761 aaaagcaggc cctgagatgg acaagagatt agcaactgaa tttgagctga gaaaacatgc
     2821 taggcaagaa ggacgcagat actgtgatcc tgttgtttta acatatcaag ctgaacggat
     2881 gccagagcaa atcaggctga aagttggtgg tgtggaccca aagcagttgg ctgtttacga
     2941 agagtttgca cgcaatgttc ctggcttctt acctacaaat gacttaagtc agcccacggg
     3001 atttttagcc cagcccatga agcaagcttg ggcaacatag ccgcgggccc gggatccacc
     3061 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3121 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3301 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3361 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3421 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3481 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3541 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3601 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3661 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3721 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3781 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3841 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3901 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4441 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4801 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5101 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5221 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5281 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5341 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5401 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5461 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5521 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6361 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     6421 gtcattagtt catagcccat atatggagtt ccg
//
