LOCUS       sequence-289255-        7104 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 7104)
  AUTHORS   Petit AP, Wohlbold L, Bawankar P, Huntzinger E, Schmidt S, 
            Izaurralde E, Weichenrieder O
  TITLE     The structural basis for the interaction between the CAF1 nuclease 
            and the NOT1 scaffold of the human CCR4-NOT deadenylase complex.
  JOURNAL   Nucleic Acids Res. 2012 Nov;40(21):11058-72. doi: 
            10.1093/nar/gks883. Epub 2012 Sep 12.
  PUBMED    22977175
REFERENCE   2  (bases 1 to 7104)
  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: 
            "10.1093/nar/gks883"; journalName: "Nucleic Acids Res"; date: 
            "2012-11"; volume: "40"; issue: "21"; pages: "11058-72"
COMMENT     Sequence Label: pT7-EGFP-C1-HsNot1iso1_1595-2376_K
FEATURES             Location/Qualifiers
     source          1..7104
                     /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             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /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(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3892..4013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3929..3948)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3983..4002
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4020..4475)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4157..4178)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4369..4388
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4502..4606
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4608..4965
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4608..4799
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4631..4651)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4816..4951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5000..5794
                     /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             5000..5794
                     /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(5054..5073)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5664..5683
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5982..6001)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6026..6073
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6402..6990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6891..6910
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctttagccca gcccatgaag caagcttggg caacagatga
     1441 tgtagctcag atttatgata agtgtattac agaactggag caacatctac atgccatccc
     1501 accaactttg gccatgaacc ctcaagctca ggctcttcga agtctcttgg aggttgtagt
     1561 tttatctcga aactctcggg atgccatagc tgctcttgga ttgctccaaa aggctgtaga
     1621 gggcttacta gatgccacaa gtggtgctga tgctgacctt ctgctgcgct acagggaatg
     1681 ccacctcttg gtcctaaaag ctctgcagga tggccgggca tatgggtctc catggtgcaa
     1741 caaacagatc acaaggtgcc taattgaatg tcgagatgaa tataaatata atgtggaggc
     1801 tgtggagctg ctaattcgca atcatttggt taatatgcag cagtatgatc ttcacctagc
     1861 gcagtcaatg gagaatggct taaactacat ggctgtggca tttgctatgc agttagtaaa
     1921 aatcctgctg gtggatgaaa ggagtgttgc tcatgttact gaggcagatc tgttccacac
     1981 cattgaaacc ctcatgagga ttaatgctca ttccagaggc aatgctccag aaggattgcc
     2041 ccagctgatg gaagtagtgc gatccaacta tgaagcaatg attgatcgtg ctcatggagg
     2101 cccaaacttt atgatgcatt ctgggatctc tcaagcctca gagtatgatg accctccagg
     2161 cctgagggag aaggcagagt atcttctgag ggaatgggtg aatctctacc attcagcagc
     2221 agctggccgc gacagtacca aagctttctc tgcatttgtt ggacagatgc accagcaagg
     2281 aatactgaag accgatgatc tcataacaag gttctttcgt ctgtgtactg aaatgtgtgt
     2341 tgaaatcagt taccgtgctc aggctgagca gcagcacaat cctgctgcca atcccaccat
     2401 gatccgagcc aagtgctatc acaacctgga tgcctttgtt cgactcattg cactgctcgt
     2461 gaaacactca ggggaggcca ccaacactgt cacaaagatt aatctgctga acaaggtcct
     2521 tggtatagta gtgggagttc tccttcagga tcatgatgtt cgtcagagtg aatttcagca
     2581 acttccctac catcgaattt ttatcatgct tctcttggaa ctcaatgcac ctgagcatgt
     2641 gttggaaacc attaatttcc agacacttac agctttctgc aatacattcc acatcttgag
     2701 gcctaccaaa gctcctggct ttgtatatgc ctggcttgaa ctgatttccc atcggatatt
     2761 tattgcaaga atgctggcac atacgccaca gcagaagggg tggcctatgt atgcacagct
     2821 actgattgat ttattcaaat atttagcgcc tttccttaga aatgtggaac tcaccaaacc
     2881 tatgcaaatc ctctacaagg gcactttaag agtgctgctg gttcttttgc atgatttccc
     2941 agagttcctt tgtgattacc attatgggtt ctgtgatgtg atcccaccta attgtatcca
     3001 gttaagaaat ttgatcctga gtgcctttcc aagaaacatg aggctccccg acccattcac
     3061 tcctaatcta aaggtggaca tgttgagtga aattaacatt gctccccgga ttctcaccaa
     3121 tttcactgga gtaatgccac ctcagttcaa aaaggatttg gattcctatc ttaaaactcg
     3181 atcaccagtc actttcctgt ctgatctgcg cagcaaccta caggtatcca atgaacctgg
     3241 gaatcgctac aacctccagc tcatcaatgc actggtgctc tatgtcggga ctcaggccat
     3301 tgcgcacatc cacaacaagg gcagcacacc ttcaatgagc accatcactc actcagcaca
     3361 catggatatc ttccagaatt tggctgtgga cttggacact gagggtcgct atctcttttt
     3421 gaatgcaatt gcaaatcagc tccggtaccc aaatagccac actcactact tcagttgcac
     3481 catgctgtac ctttttgcag aggccaatac ggaagccatc caagaacaga tcacaagagt
     3541 tctcttggaa cggttgattg taaataggcc acatccttgg ggtcttctta ttaccttcat
     3601 tgagctgatt aaaaacccag cgtttaagtt ctggaaccat gaatttgtac actgtgcccc
     3661 agaaatcgaa aagttattcc agtcggtcgc acagtgctgc atgggacaga agcaggccca
     3721 gcaagtaatg gaagggacag gtgccagtta gccgcgggcc cgggatccac cggatctaga
     3781 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3841 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3901 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3961 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     4021 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4081 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4141 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4201 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4261 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4321 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4381 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4441 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4501 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4561 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4621 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4681 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4741 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4801 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4861 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4921 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4981 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5041 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5101 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5161 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5221 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5281 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5341 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5401 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5461 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5521 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5581 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5641 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5701 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5761 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5821 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5881 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5941 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     6001 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     6061 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     6121 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     6181 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     6241 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6301 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6361 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6421 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6481 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6541 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6601 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6661 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6721 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6781 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6841 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6901 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6961 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7021 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7081 gataaccgta ttaccgccat gcat
//
