LOCUS       sequence-289938-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1595-2376.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  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 7236)
  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-EGFP-C1-HsNot1iso1_1595-2376_S
FEATURES             Location/Qualifiers
     source          1..7236
                     /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"
     CDS             join(797..799,800..802,803..1513)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(797..799,800..802,803..1513)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(842..863)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1103..1122)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1450..1471
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4024..4145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4061..4080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4115..4134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4152..4607)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4289..4310)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4501..4520
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4634..4738
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4740..5097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4740..4931
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4763..4783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4948..5083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5132..5926
                     /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             5132..5926
                     /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(5186..5205)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5796..5815
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6114..6133)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6158..6205
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6534..7122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7023..7042
                     /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 cgacctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      841 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      901 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      961 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1021 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1081 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1141 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1201 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1261 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1321 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1381 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1441 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1501 cgagctgtac aagtccggac tcagatctcg agctttagcc cagcccatga agcaagcttg
     1561 ggcaacagat gatgtagctc agatttatga taagtgtatt acagaactgg agcaacatct
     1621 acatgccatc ccaccaactt tggccatgaa ccctcaagct caggctcttc gaagtctctt
     1681 ggaggttgta gttttatctc gaaactctcg ggatgccata gctgctcttg gattgctcca
     1741 aaaggctgta gagggcttac tagatgccac aagtggtgct gatgctgacc ttctgctgcg
     1801 ctacagggaa tgccacctct tggtcctaaa agctctgcag gatggccggg catatgggtc
     1861 tccatggtgc aacaaacaga tcacaaggtg cctaattgaa tgtcgagatg aatataaata
     1921 taatgtggag gctgtggagc tgctaattcg caatcatttg gttaatatgc agcagtatga
     1981 tcttcaccta gcgcagtcaa tggagaatgg cttaaactac atggctgtgg catttgctat
     2041 gcagttagta aaaatcctgc tggtggatga aaggagtgtt gctcatgtta ctgaggcaga
     2101 tctgttccac accattgaaa ccctcatgag gattaatgct cattccagag gcaatgctcc
     2161 agaaggattg ccccagctga tggaagtagt gcgatccaac tatgaagcaa tgattgatcg
     2221 tgctcatgga ggcccaaact ttatgatgca ttctgggatc tctcaagcct cagagtatga
     2281 tgaccctcca ggcctgaggg agaaggcaga gtatcttctg agggaatggg tgaatctcta
     2341 ccattcagca gcagctggcc gcgacagtac caaagctttc tctgcatttg ttggacagat
     2401 gcaccagcaa ggaatactga agaccgatga tctcataaca aggttctttc gtctgtgtac
     2461 tgaaatgtgt gttgaaatca gttaccgtgc tcaggctgag cagcagcaca atcctgctgc
     2521 caatcccacc atgatccgag ccaagtgcta tcacaacctg gatgcctttg ttcgactcat
     2581 tgcactgctc gtgaaacact caggggaggc caccaacact gtcacaaaga ttaatctgct
     2641 gaacaaggtc cttggtatag tagtgggagt tctccttcag gatcatgatg ttcgtcagag
     2701 tgaatttcag caacttccct accatcgaat ttttatcatg cttctcttgg aactcaatgc
     2761 acctgagcat gtgttggaaa ccattaattt ccagacactt acagctttct gcaatacatt
     2821 ccacatcttg aggcctacca aagctcctgg ctttgtatat gcctggcttg aactgatttc
     2881 ccatcggata tttattgcaa gaatgctggc acatacgcca cagcagaagg ggtggcctat
     2941 gtatgcacag ctactgattg atttattcaa atatttagcg cctttcctta gaaatgtgga
     3001 actcaccaaa cctatgcaaa tcctctacaa gggcacttta agagtgctgc tggttctttt
     3061 gcatgatttc ccagagttcc tttgtgatta ccattatggg ttctgtgatg tgatcccacc
     3121 taattgtatc cagttaagaa atttgatcct gagtgccttt ccaagaaaca tgaggctccc
     3181 cgacccattc actcctaatc taaaggtgga catgttgagt gaaattaaca ttgctccccg
     3241 gattctcacc aatttcactg gagtaatgcc acctcagttc aaaaaggatt tggattccta
     3301 tcttaaaact cgatcaccag tcactttcct gtctgatctg cgcagcaacc tacaggtatc
     3361 caatgaacct gggaatcgct acaacctcca gctcatcaat gcactggtgc tctatgtcgg
     3421 gactcaggcc attgcgcaca tccacaacaa gggcagcaca ccttcaatga gcaccatcac
     3481 tcactcagca cacatggata tcttccagaa tttggctgtg gacttggaca ctgagggtcg
     3541 ctatctcttt ttgaatgcaa ttgcaaatca gctccggtac ccaaatagcc acactcacta
     3601 cttcagttgc accatgctgt acctttttgc agaggccaat acggaagcca tccaagaaca
     3661 gatcacaaga gttctcttgg aacggttgat tgtaaatagg ccacatcctt ggggtcttct
     3721 tattaccttc attgagctga ttaaaaaccc agcgtttaag ttctggaacc atgaatttgt
     3781 acactgtgcc ccagaaatcg aaaagttatt ccagtcggtc gcacagtgct gcatgggaca
     3841 gaagcaggcc cagcaagtaa tggaagggac aggtgccagt tagccgcggg cccgggatcc
     3901 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3961 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4021 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4081 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4141 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4201 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4261 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4321 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4381 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4441 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4501 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4561 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4741 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4801 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4861 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4921 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4981 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5041 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     5101 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5161 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5221 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5281 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     5341 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5401 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5461 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5521 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5581 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5641 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5701 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5761 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5821 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5881 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5941 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6001 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6061 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     6121 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     6181 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     6241 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     6301 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     6361 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     6421 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6481 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6541 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6601 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6661 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6721 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6781 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6841 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6901 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6961 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7021 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7081 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7141 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7201 cctgattctg tggataaccg tattaccgcc atgcat
//
