LOCUS       sequence-289930-        7555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  AUTHORS   Boland A, Chen Y, Raisch T, Jonas S, Kuzuoglu-Ozturk D, Wohlbold L, 
            Weichenrieder O, Izaurralde E
  TITLE     Structure and assembly of the NOT module of the human CCR4-NOT 
            complex.
  JOURNAL   Nat Struct Mol Biol. 2013 Nov;20(11):1289-97. doi: 
            10.1038/nsmb.2681. Epub 2013 Oct 13.
  PUBMED    24121232
REFERENCE   2  (bases 1 to 7555)
  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: "10.1038/nsmb.2681";
            journalName: "Nat Struct Mol Biol"; date: "2013-11"; volume: "20"; 
            issue: "11"; pages: "1289-97"
COMMENT     Sequence Label: pCIneo-lambdaN-HsNot9-HA-MBP_S
FEATURES             Location/Qualifiers
     source          1..7555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..1160
                     /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             2091..2117
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3075..3084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3259..3380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3270..3289)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3324..3343
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3566..4021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3653..3672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3863..3884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4138..4495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4138..4329
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4161..4181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4346..4481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4546..5340
                     /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(4600..4619)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5210..5229
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5404..5452
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5493..5512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5612..5634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5672..5690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5758..5862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5863..5931,5932..6723)
                     /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     complement(6081..6100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6894..7482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7383..7402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag
     1141 ctcaatggaa agctgcaaac ccaccgctcg agaattcaat gcacagcctg gcgacggctg
     1201 cgcctgtgcc tactacactg gcacaagtgg atagagaaaa gatctatcag tggatcaatg
     1261 agctgtccag tcctgagact agggaaaatg ctttgctgga gctaagtaag aagcgagaat
     1321 ctgttcctga ccttgcaccc atgctgtggc attcatttgg tactattgca gcacttttac
     1381 aggaaattgt aaatatttat ccatctatca acccacccac cttgacagca caccagtcta
     1441 acagagtttg caatgctctg gcattactgc aatgtgtagc atcacatcca gaaaccaggt
     1501 cagcgtttct cgcagcacac atcccacttt ttttgtaccc ctttttgcac actgtcagca
     1561 aaacacgtcc ctttgagtat ctccggctca ccagccttgg agttattggg gccctggtga
     1621 aaacagatga acaagaagta atcaactttt tattaacaac agaaattatc cctttatgtt
     1681 tgcgaattat ggaatctgga agtgaacttt ctaaaacagt tgccacattc atcctccaga
     1741 agatcttgtt agatgacact ggtttggctt atatatgtca gacgtatgag cgtttctccc
     1801 atgttgccat gatcttgggt aagatggtcc tgcagctatc caaagagcct tctgcccgtc
     1861 tgctgaagca tgtagtgaga tgttaccttc gactttcaga taaccccagg gcacgtgaag
     1921 cactcagaca gtgcctccct gaccagctga aagacacaac cttcgcccag gtgctaaaag
     1981 atgacaccac cacgaaacgc tggcttgcac aactggtgaa gaacctgcaa gagggccagg
     2041 tcaccgatcc ccggggtatc cccctgcccc ctcagaagaa ttcaatggct tacccatacg
     2101 atgttccaga ttacgctatg aaaatcgaag aaggtaaact ggtaatctgg attaacggcg
     2161 ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat accggaatta
     2221 aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccgcaggtt gcggcaactg
     2281 gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac gctcaatctg
     2341 gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat ccgtttacct
     2401 gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat
     2461 cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa gagatcccgg
     2521 cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac
     2581 cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag tatgaaaacg
     2641 gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct
     2701 tcctggttga cctgattaaa agcaaacaca tgaatgcaga caccgattac tccatcgcag
     2761 aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca
     2821 acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc aagggtcaac
     2881 catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt ccgaacaaag
     2941 agctggcgaa agagttcctc gaaaactatc tgctgactga tgaaggtctg gaagcggtta
     3001 ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag
     3061 atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc
     3121 cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc
     3181 gtcagactgt cgatgaagcc ctgaaagatt aggcggccgc ttccctttag tgagggttaa
     3241 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3301 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3361 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3421 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     3481 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3541 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3781 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3901 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3961 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4021 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     4081 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     4141 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4201 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4321 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4381 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4441 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4501 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     4561 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4621 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4681 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4741 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4801 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4861 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4921 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4981 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5041 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5101 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5161 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5221 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5281 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5341 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     5401 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5461 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5521 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5581 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5641 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5701 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5881 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5941 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6001 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6061 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6121 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6181 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6241 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6301 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6361 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6421 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6481 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6541 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6601 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6661 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6721 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6781 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6841 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6901 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6961 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7021 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7081 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7141 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7201 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7261 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7321 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7381 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7441 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7501 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
