LOCUS       sequence-291207-        7515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot4-del377-402.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7515)
  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 7515)
  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-del377-402_AG
FEATURES             Location/Qualifiers
     source          1..7515
                     /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(3071..3089)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3072..3089)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3109..3126)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3115..3339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3385..3813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3472..3491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3682..3703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3822..3842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3827..4156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4007..4142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4069..4088
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4223..5017
                     /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(4277..4296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4887..4906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5191..5312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5228..5247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5282..5301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5361..5377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5361..5377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5374..5396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5385..5401
                     /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(5409..5415,5416..5433,5434..5439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5454..5475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5592..5609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5763..6348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5843..5862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6519..7310,7311..7379))
                     /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     7142..7161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7380..7484)
                     /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 gtatgatacc
     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 acatcattgg gttttgatcc cttcgatgtc actcgaaaag ccttagcaga cctgattgag
     2581 aaggaactgt ccgttcaaga ccaaccttcc ctttcgccca catctcttca gaactcctct
     2641 tcacacacta caaccgccaa aggtccaggc tctggattcc tgcatcctgc tgcagctaca
     2701 aatgccaatt ctctcaatag taccttttca gtcttgcccc agaggttccc tcaatttcag
     2761 cagcaccgag cggtttataa ttcattcagt tttccaggcc aggcagcccg ctatccttgg
     2821 atggcctttc cacgcaatag catcatgcac ttgaaccaca cagcaaaccc cacctcaaat
     2881 agtaatttct tggacttgaa tctcccgcca cagcacaaca caggtctggg agggatccct
     2941 gtagcagggg aagaagaggt gaaggtttcg accatgccac tgtcaacctc ttcccattca
     3001 ttacaacaag gacagcagcc tacaagtctc cacactactg tggcctaagc ggccgcctag
     3061 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     3121 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3181 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3241 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3301 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3361 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3421 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3481 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3541 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3601 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3661 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3721 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3781 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3901 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3961 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4021 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4081 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4141 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4201 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4261 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4321 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4381 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4441 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4501 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4561 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4621 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4681 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4741 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4801 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4861 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4921 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4981 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5041 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5101 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5161 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5341 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5401 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5461 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5521 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6181 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6361 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6481 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6541 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6601 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6661 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6721 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6781 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6841 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6901 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6961 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7021 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7081 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7141 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7201 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7261 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7321 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7441 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7501 gtgccacctg acgtc
//
