LOCUS       sequence-290074-        6364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot3-del632-654.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  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 6364)
  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: pLNHA-C1-HsNot3-del632-654_T
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             665..730
                     /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             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3152..3273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3189..3208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3243..3262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3280..3735)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3417..3438)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3629..3648
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3762..3866
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3868..4225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3868..4059
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3891..3911)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4076..4211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4260..5054
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4260..5054
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4314..4333)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4924..4943
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5242..5261)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5286..5333
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5662..6250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6151..6170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcgcggacaa
      841 gcgcaaactc caaggtgaga ttgatcgctg cctcaagaag gtgtccgagg gcgtggagca
      901 gtttgaagat atttggcaga agctccacaa tgcagccaac gcgaaccaga aagaaaagta
      961 tgaggctgac ctaaagaagg agattaagaa gctacaacgg ctgagggacc aaatcaagac
     1021 atgggtagcg tccaacgaga tcaaggacaa gaggcagctt atagacaacc gcaagctcat
     1081 tgagacgcaa atggaacggt tcaaagttgt ggaacgagag accaaaacca aagcttacag
     1141 caaagagggc ctgggcctgg cccagaaggt agatcctgcc cagaaggaga aggaagaggt
     1201 tggccagtgg ctcacgaata ccatcgacac gctcaacatg caggtggacc agtttgagag
     1261 tgaagtggag tcactgtcag tgcagacacg caagaagaag ggcgacaagg ataagcagga
     1321 ccggattgag ggcttgaagc ggcacatcga gaagcaccgc taccacgtgc gcatgctaga
     1381 gaccatcctg cgcatgctgg acaatgactc catcctcgtt gacgccatcc gcaagatcaa
     1441 ggacgacgtt gagtactatg ttgactcatc ccaggacccc gacttcgagg agaacgagtt
     1501 tctctacgat gacctggacc tcgaggacat tccacaggcg ctggtcgcca cctccccccc
     1561 cagccacagc cacatggagg atgagatctt caaccagtcc agcagcacgc ccacctcaac
     1621 cacctccagc tctcccatcc cgcccagccc agccaactgt accacggaaa actctgaaga
     1681 tgataagaag aggggacgtt ccacagacag tgaagtcagc cagtctccag ccaaaaacgg
     1741 ctccaagcct gtccacagca accagcaccc tcagtcccca gctgtgccgc ccacctaccc
     1801 ctccggcccc ccgcctgctg cctctgcctt gagcaccact cctggcaaca atggggtccc
     1861 cgcccccgca gcacccccaa gtgccctggg ccccaaggcc agtccagctc ccagccacaa
     1921 ctcgggcacc cctgctccct atgcccaggc tgtggcccca ccagctccca gtgggcccag
     1981 cacgacccag ccccggcccc ccagcgtcca gcctagcgga ggcggaggcg gcggcagcgg
     2041 aggtggaggg agcagcagca gtagtaacag cagtgccggt ggaggggctg gcaagcagaa
     2101 tggcgccacc agttacagct cagttgtggc agacagcccg gcagaggtgg ctttgagcag
     2161 cagtgggggc aacaatgcca gcagccaggc cttgggcccc ccttccggcc cccacaaccc
     2221 acctcccagc acctcgaagg aacccagtgc ggcagcccca acgggggctg ggggcgtggc
     2281 cccaggctca gggaacaact cagggggacc cagcctcctg gtgccactgc ctgtgaatcc
     2341 tcccagctcc ccaacgccca gcttcagtga tgccaaggca gccggtgccc tgctcaatgg
     2401 gcctccacag ttcagcaccg ccccagaaat caaggcccct gagcctctga gctccttgaa
     2461 gtccatggcg gaacgggcag ccatcagctc tggcattgag gaccctgtgc caacgctgca
     2521 cctgaccgag cgagacatca tcctgagcag tacatcagca cctccggcct cagcccagcc
     2581 gcccctgcag ctgtcagagg tgaacatacc gctgtcgctg ggtgtctgtc cactgggccc
     2641 tgtgcccctc accaaggagc agctctatca gcaggccatg gaagaggccg cctggcacca
     2701 catgcctcac ccctctgact ctccacactc ggacactgtg gaattctacc agcgcctgtc
     2761 gaccgagaca ctcttcttca tcttctacta tctggagggc actaaggcac agtatctggc
     2821 agccaaggcc ctaaagaagc agtcatggcg attccacacc aagtacatga tgtggttcca
     2881 gaggcacgag gagcccaaga ccatcactga cgagtttgag cagggcacct acatctactt
     2941 tgactacgag aagtggggcc agcggaagaa ggaaggcttc acctttgagt accgctacct
     3001 ggaggaccgg gacctccagt gaggatccac cggatctaga taactgatca taatcagcca
     3061 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3121 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3181 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3241 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     3301 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3361 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3421 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3481 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3541 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3601 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3661 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3721 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3781 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3841 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3901 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3961 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4021 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4081 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4141 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4201 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4261 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4321 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4381 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4441 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4501 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4561 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4621 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4681 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4741 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4801 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4861 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4921 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4981 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5041 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5101 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5161 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5221 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5281 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5341 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5401 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5461 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5521 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5581 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5641 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5701 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5761 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5821 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5881 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5941 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6001 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6061 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6361 gcat
//
