LOCUS       sequence-291208-        7449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot4-del377-424.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7449)
  AUTHORS   Keskeny C, Raisch T, Sgromo A, Igreja C, Bhandari D, Weichenrieder 
            O, Izaurralde E
  TITLE     A conserved CAF40-binding motif in metazoan NOT4 mediates 
            association with the CCR4-NOT complex.
  JOURNAL   Genes Dev. 2019 Feb 1;33(3-4):236-252. doi: 10.1101/gad.320952.118. 
            Epub 2019 Jan 28.
  PUBMED    30692204
REFERENCE   2  (bases 1 to 7449)
  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"; journalName: "Genes 
            Dev."; date: "2019-02-1"; volume: "33(3-4):236-252. doi"; pages: " 
            10.1101/gad.320952.118. Epub 2019 Jan 28"
COMMENT     Sequence Label: pcDNA3.1-MS2-HA-HsNot4-del377-424_AG
FEATURES             Location/Qualifiers
     source          1..7449
                     /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(3005..3023)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3006..3023)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3043..3060)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3049..3273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3319..3747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3406..3425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3616..3637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3756..3776)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3761..4090
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3941..4076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4003..4022
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4157..4951
                     /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(4211..4230)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4821..4840
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5125..5246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5162..5181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5216..5235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5295..5311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5295..5311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5308..5330)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5319..5335
                     /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(5343..5349,5350..5367,5368..5373))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5388..5409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5526..5543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5697..6282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5777..5796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6453..7244,7245..7313))
                     /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     7076..7095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7314..7418)
                     /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 gactcgagat gtctcgcagt cctgatgcga aggaagaccc tgtggagtgc
     1441 cctctttgca tggagccctt ggagatagat gatatcaact ttttcccttg cacctgtggc
     1501 taccagattt gccgattttg ttggcatcga attcgcactg atgaaaatgg gctttgtcct
     1561 gcatgtagaa agccatatcc agaagaccca gcagtttata aaccactctc ccaggaagag
     1621 ctgcaaagga taaagaatga gaaaaaacag aaacaaaatg agagaaaaca gaaaatatca
     1681 gaaaatcgca aacatttggc tagtgtacgt gtcgtacaaa aaaacctcgt ctttgttgta
     1741 ggtttatctc agcgcctagc agacccagag gttttaaaac gaccagaata ttttgggaag
     1801 tttggtaaaa tacataaagt tgtcatcaat aatagcacat catatgcagg ctcacagggt
     1861 ccaagtgcca gtgcttatgt aacctatatc cggtcagaag acgctctcag agccatacag
     1921 tgtgtcaaca atgtggtagt agatggcaga acacttaagg catctctagg tacaacaaaa
     1981 tactgcagtt acttcttaaa gaatatgcag tgtccaaaac ctgactgcat gtatcttcat
     2041 gaattggggg atgaggcggc cagcttcaca aaagaggaaa tgcaggcggg taaacaccaa
     2101 gaatatgaac agaagctact tcaagaatta tataaattaa atcccaattt tcttcagcta
     2161 tctacgggtt cagttgataa aaataagaac aaagtgacac cactgcagag gtacgatacc
     2221 cccattgaca aaccttcaga ttctctcagt atagggaacg gtgataattc ccagcagata
     2281 tctaacagtg atacgccttc accaccacct ggtttgtcaa aatccaatcc agtcatcccc
     2341 atcagttcat ccaatcacag tgcacggtcc ccttttgaag gggcagtaac agagtcacag
     2401 tcgttattct cagacaattt tcgccatccc aaccctatcc caagtgggct tcctcctttc
     2461 cccagctccc cacagacatc cagtgactgg cctacagcac cagaaccaca gagcctcttc
     2521 acatcagttc aagaccaacc ttccctttcg cccacatctc ttcagaactc ctcttcacac
     2581 actacaaccg ccaaaggtcc aggctctgga ttcctgcatc ctgctgcagc tacaaatgcc
     2641 aattctctca atagtacctt ttcagtcttg ccccagaggt tccctcaatt tcagcagcac
     2701 cgagcggttt ataattcatt cagttttcca ggccaggcag cccgctatcc ttggatggcc
     2761 tttccacgca atagcatcat gcacttgaac cacacagcaa accccacctc aaatagtaat
     2821 ttcttggact tgaatctccc gccacagcac aacacaggtc tgggagggat ccctgtagca
     2881 ggggaagaag aggtgaaggt ttcgaccatg ccactgtcaa cctcttccca ttcattacaa
     2941 caaggacagc agcctacaag tctccacact actgtggcct aagcggccgc ctagagggcc
     3001 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     3061 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3121 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3181 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3241 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3301 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3361 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3421 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3481 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3541 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3601 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3661 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3721 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3781 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3841 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3961 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4021 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4081 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4141 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4201 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4261 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4321 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4381 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4441 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4501 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4561 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4621 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4681 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4741 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4801 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4861 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4921 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4981 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5041 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5101 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5161 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5221 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5281 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5341 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5401 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5461 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5521 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5581 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6121 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     6241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6481 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6541 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6601 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6661 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6721 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6781 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6841 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6901 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6961 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7441 cctgacgtc
//
