LOCUS       sequence-290448-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_840-1605-IKGRQtoDSYAA.
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-IKGRQtoDSYAA_X
FEATURES             Location/Qualifiers
     source          1..6453
                     /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    3241..3362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3278..3297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3332..3351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3369..3824)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3506..3527)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3718..3737
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3851..3955
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3957..4314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3957..4148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3980..4000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4165..4300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4349..5143
                     /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             4349..5143
                     /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(4403..4422)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5013..5032
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5331..5350)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5375..5422
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5751..6339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6240..6259
                     /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 gagaaaattg cttttatttt caataatctc tcacagtcaa atatgacaca
     1621 aaaggttgaa gagctaaagg aaacggtgaa agaagaattt atgccttggg tttcacagta
     1681 tctggttatg aagagagtca gtattgagcc aaactttcat agcctgtatt caaacttcct
     1741 tgacacgctg aagaatcctg aatttaacaa gatggttctg aatgagacct acagaaacat
     1801 taaagtgctc ctgacctctg ataaagctgc agccaatttc tcagatcgtt ctttgctgaa
     1861 gaacttggga cattggctag gaatgatcac attagctaaa aacaaaccca tcttacacac
     1921 tgacttggat gtgaaatcat tgctgctaga ggcttatgtt aaaggacaac aagaattgct
     1981 ctatgtagtg ccctttgttg ccaaagtctt agaatctagc attcgtagtg tggtttttag
     2041 gccaccaaac ccttggacaa tggcaattat gaatgtatta gctgagctac atcaggagca
     2101 tgacttaaag ttaaacttga agtttgaaat cgaggttctc tgcaagaacc ttgcattaga
     2161 catcaatgag ctaaaacctg gaaacctcct aaaggataaa gatcgcctga agaatttaga
     2221 tgagcaactc tctgctccaa agaaagatgt caagcagcca gaagaactcc ctcccatcac
     2281 aaccacaaca acttctacta caccagctac caacaccact tgtacagcca cggttccacc
     2341 acagccacag tacagctacc acgacatcaa tgtctattcc cttgcgggct tggcaccaca
     2401 cattactcta aatccaacaa ttcccttgtt tcaggcccat ccacagttga agcagtgtgt
     2461 gcgtcaggca attgaacggg ctgtccagga gctggtccat cctgtggtgg atcgatcaat
     2521 taagattgcc atgactactt gtgagcaaga tgtcaggtcg gattttgccc tggattcgga
     2581 ggaatctcga atgcgaatag cagctcatca catgatgcgt aacttgacag cttacatggc
     2641 tatgattaca tgcgctgaac ctttgctcat gagcatatct accaacttaa aaaacagttt
     2701 tgcctcagcc cttcgtactg cttccccaca acaaagagaa atgatggatc aggcagctgc
     2761 tcaattagct caggacaatt gtgagttggc ttgctgtttt attcagaaga ctgcagtaga
     2821 aaaagcaggc cctgagatgg acaagagatt agcaactgaa tttgagctga gaaaacatgc
     2881 taggcaagaa ggacgcagat actgtgatcc tgttgtttta acatatgctg ctgaacggat
     2941 gccagagcaa atcaggctga aagttggtgg tgtggaccca aagcagttgg ctgtttacga
     3001 agagtttgca cgcaatgttc ctggcttctt acctacaaat gacttaagtc agcccacggg
     3061 atttttagcc cagcccatga agcaagcttg ggcaacatag ccgcgggccc gggatccacc
     3121 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3181 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3241 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3301 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3361 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3481 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3541 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3601 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3661 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3721 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3781 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3841 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3901 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3961 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4201 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4261 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4321 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4381 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4441 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4501 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4561 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4621 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4681 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4741 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4801 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4861 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4921 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4981 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5041 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5101 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5161 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5221 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5281 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5341 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5401 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5461 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5521 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5581 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5881 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6421 gattctgtgg ataaccgtat taccgccatg cat
//
