LOCUS       sequence-290864-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1841-2376-shRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Kuzuoglu-Ozturk D, Bhandari D, Huntzinger E, Fauser M, Helms S, 
            Izaurralde E
  TITLE     miRISC and the CCR4-NOT complex silence mRNA targets independently 
            of 43S ribosomal scanning.
  JOURNAL   EMBO J. 2016 Jun 1;35(11):1186-203. doi: 10.15252/embj.201592901. 
            Epub 2016 Mar 23.
  PUBMED    27009120
REFERENCE   2  (bases 1 to 7487)
  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: 
            "10.15252/embj.201592901"; journalName: "EMBO J"; date: "2016-06-1- 
            1"; volume: "35"; issue: "11"; pages: "1186-203"
COMMENT     Sequence Label: pcDNA3.1-MS2-HA-HsNot1iso1_1841-2376-shRNAres_AC
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3043..3061)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3044..3061)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3081..3098)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3087..3311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3357..3785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3444..3463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3654..3675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3794..3814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3799..4128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3979..4114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4041..4060
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4195..4989
                     /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(4249..4268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4859..4878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5163..5284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5200..5219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5254..5273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5333..5349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5333..5349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5346..5368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5357..5373
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5381..5387,5388..5405,5406..5411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5426..5447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5564..5581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5735..6320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5815..5834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6491..7282,7283..7351))
                     /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     7114..7133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7352..7456)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccgaa
     1381 ttcccgggtc gactcgagga gtatgatgac cctccaggcc tgagggagaa ggcagagtat
     1441 cttctgaggg aatgggtgaa tctctaccat tcagcagcag ctggccgcga cagtaccaaa
     1501 gctttctctg catttgttgg acagatgcac cagcaaggaa tactgaagac cgatgatctc
     1561 ataacaaggt tctttcgtct gtgtactgaa atgtgtgttg aaatcagtta ccgtgctcag
     1621 gctgagcagc agcacaatcc tgctgccaat cccaccatga tccgagccaa gtgctatcac
     1681 aacctggatg cctttgttcg actcattgca ctgctcgtga aacactcagg ggaggccacc
     1741 aacactgtca caaagattaa tctgctgaac aaggtccttg gtatagtagt gggagttctc
     1801 cttcaggatc atgatgttcg tcagagtgaa tttcagcaac ttccctacca tcgaattttt
     1861 atcatgcttc tcttggaact caatgcacct gagcatgtgt tggaaaccat taatttccag
     1921 acacttacag ctttctgcaa tacattccac atcttgaggc ctaccaaagc tcctggcttt
     1981 gtatatgcct ggcttgaact gatttcccat cggatattta ttgcaagaat gctggcacat
     2041 acgccacagc agaaggggtg gcctatgtat gcacagctac tgattgattt attcaaatat
     2101 ttagcgcctt tccttagaaa tgtggaactc accaaaccta tgcaaatcct ctacaagggc
     2161 actttaagag tgctgctggt tcttttgcat gatttcccag agttcctttg tgattaccat
     2221 tatgggttct gtgatgtgat cccacctaat tgtatccagt taagaaattt gatcctgagt
     2281 gcctttccaa gaaacatgag gctccccgac ccattcactc ctaatctaaa ggtggacatg
     2341 ttgagtgaaa ttaacattgc tccccggatt ctcaccaatt tcactggagt aatgccacct
     2401 cagttcaaaa aggatttgga ttcctatctt aaaactcgat caccagtcac tttcctgtct
     2461 gatctgcgca gcaacctaca ggtatccaat gaacctggga atcgctacaa cctccagctc
     2521 atcaatgcac tggtgctcta tgtcgggact caggccattg cgcacatcca caacaagggc
     2581 agcacacctt caatgagcac catcactcac tcagcacaca tggatatctt ccagaatttg
     2641 gctgtggact tggacactga gggtcgctat ctctttttga atgcaattgc aaatcagctc
     2701 cggtacccaa atagccacac tcactacttc agttgcacca tgctgtacct ttttgcagag
     2761 gccaatacgg aagccatcca agaacagatc acaagagttc tcttggaacg gttgattgta
     2821 aataggccac atccttgggg tcttcttatt accttcattg agctgattaa aaacccagcg
     2881 tttaagttct ggaaccatga atttgtacac tgtgccccag aaatcgaaaa gttattccag
     2941 tcggtcgcac agtgctgcat gggacagaag caggcccagc aagtaatgga agggacaggt
     3001 gccagttagg cggccgctag gcggccgcct agagggccct attctatagt gtcacctaaa
     3061 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3121 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3181 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3241 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3301 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3421 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3541 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3781 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4081 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4141 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4201 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4261 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4321 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4381 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4441 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4501 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4561 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4621 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4681 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4741 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4801 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4861 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4921 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4981 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5041 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5101 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5161 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5221 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5281 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5341 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5401 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5461 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5521 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5581 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5641 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5701 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5761 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5821 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5881 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5941 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6001 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6061 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6121 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6181 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6241 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
