LOCUS       sequence-291210-        7593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot4-FFTLtoDAEE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7593)
  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 7593)
  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-FFTLtoDAEE_AG
FEATURES             Location/Qualifiers
     source          1..7593
                     /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(3149..3167)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3150..3167)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3187..3204)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3193..3417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3463..3891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3760..3781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3905..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4147..4166
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4301..5095
                     /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(4355..4374)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4965..4984
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5269..5390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5306..5325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5360..5379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5439..5455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5439..5455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5452..5474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5463..5479
                     /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(5487..5493,5494..5511,5512..5517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5532..5553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5670..5687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5841..6426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5921..5940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6597..7388,7389..7457))
                     /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     7220..7239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7458..7562)
                     /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 acatcagaaa caatcccagt atcatcctct acagactggc aagcagcttt tggctttggt
     2581 tcttctaaac aaccagagga tgacttgggt gatgatcccg ccgatgtcga gcgaaaagcc
     2641 gaggcagacc tgattgagaa ggaactgtcc gttcaagacc aaccttccct ttcgcccaca
     2701 tctcttcaga actcctcttc acacactaca accgccaaag gtccaggctc tggattcctg
     2761 catcctgctg cagctacaaa tgccaattct ctcaatagta ccttttcagt cttgccccag
     2821 aggttccctc aatttcagca gcaccgagcg gtttataatt cattcagttt tccaggccag
     2881 gcagcccgct atccttggat ggcctttcca cgcaatagca tcatgcactt gaaccacaca
     2941 gcaaacccca cctcaaatag taatttcttg gacttgaatc tcccgccaca gcacaacaca
     3001 ggtctgggag ggatccctgt agcaggggaa gaagaggtga aggtttcgac catgccactg
     3061 tcaacctctt cccattcatt acaacaagga cagcagccta caagtctcca cactactgtg
     3121 gcctaagcgg ccgcctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     3181 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3241 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3301 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3361 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3421 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4261 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4321 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4381 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4441 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4501 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4561 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4621 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4681 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4741 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4801 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4861 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4921 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4981 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5041 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5101 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5161 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5221 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5281 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5341 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5401 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5461 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5521 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5581 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5641 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5701 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5761 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6001 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6361 accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6481 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6541 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6601 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6661 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6721 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6781 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6841 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6901 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6961 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7021 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7081 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7141 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7201 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7261 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7321 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7381 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7441 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7501 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7561 cgcacatttc cccgaaaagt gccacctgac gtc
//
