LOCUS       sequence-289254-        6321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1085-1605.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6321)
  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 6321)
  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_1085-1605_K
FEATURES             Location/Qualifiers
     source          1..6321
                     /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    3109..3230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3146..3165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3200..3219
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3237..3692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3374..3395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3586..3605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3719..3823
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3825..4182
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3825..4016
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3848..3868)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4033..4168
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4217..5011
                     /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             4217..5011
                     /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(4271..4290)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4881..4900
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5199..5218)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5243..5290
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5619..6207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6108..6127
                     /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 ctactgagag aattgtggag cccccagaaa atatccagga
     1441 gaaaattgct tttattttca ataatctctc acagtcaaat atgacacaaa aggttgaaga
     1501 gctaaaggaa acggtgaaag aagaatttat gccttgggtt tcacagtatc tggttatgaa
     1561 gagagtcagt attgagccaa actttcatag cctgtattca aacttccttg acacgctgaa
     1621 gaatcctgaa tttaacaaga tggttctgaa tgagacctac agaaacatta aagtgctcct
     1681 gacctctgat aaagctgcag ccaatttctc agatcgttct ttgctgaaga acttgggaca
     1741 ttggctagga atgatcacat tagctaaaaa caaacccatc ttacacactg acttggatgt
     1801 gaaatcattg ctgctagagg cttatgttaa aggacaacaa gaattgctct atgtagtgcc
     1861 ctttgttgcc aaagtcttag aatctagcat tcgtagtgtg gtttttaggc caccaaaccc
     1921 ttggacaatg gcaattatga atgtattagc tgagctacat caggagcatg acttaaagtt
     1981 aaacttgaag tttgaaatcg aggttctctg caagaacctt gcattagaca tcaatgagct
     2041 aaaacctgga aacctcctaa aggataaaga tcgcctgaag aatttagatg agcaactctc
     2101 tgctccaaag aaagatgtca agcagccaga agaactccct cccatcacaa ccacaacaac
     2161 ttctactaca ccagctacca acaccacttg tacagccacg gttccaccac agccacagta
     2221 cagctaccac gacatcaatg tctattccct tgcgggcttg gcaccacaca ttactctaaa
     2281 tccaacaatt cccttgtttc aggcccatcc acagttgaag cagtgtgtgc gtcaggcaat
     2341 tgaacgggct gtccaggagc tggtccatcc tgtggtggat cgatcaatta agattgccat
     2401 gactacttgt gagcaaatag tcaggaagga ttttgccctg gattcggagg aatctcgaat
     2461 gcgaatagca gctcatcaca tgatgcgtaa cttgacagct ggaatggcta tgattacatg
     2521 cagggaacct ttgctcatga gcatatctac caacttaaaa aacagttttg cctcagccct
     2581 tcgtactgct tccccacaac aaagagaaat gatggatcag gcagctgctc aattagctca
     2641 ggacaattgt gagttggctt gctgttttat tcagaagact gcagtagaaa aagcaggccc
     2701 tgagatggac aagagattag caactgaatt tgagctgaga aaacatgcta ggcaagaagg
     2761 acgcagatac tgtgatcctg ttgttttaac atatcaagct gaacggatgc cagagcaaat
     2821 caggctgaaa gttggtggtg tggacccaaa gcagttggct gtttacgaag agtttgcacg
     2881 caatgttcct ggcttcttac ctacaaatga cttaagtcag cccacgggat ttttagccca
     2941 gcccatgaag caagcttggg caacatgacc gcgggcccgg gatccaccgg atctagataa
     3001 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3061 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3121 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3181 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     3241 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3301 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3361 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3421 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3481 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3541 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3601 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3661 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3721 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3781 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3841 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4021 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4081 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4141 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4201 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4261 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4321 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4381 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4441 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4501 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4561 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4621 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4681 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4741 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4801 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4861 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4921 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4981 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5041 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5101 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5161 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5221 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5281 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5341 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5401 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5461 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5521 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5761 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6301 aaccgtatta ccgccatgca t
//
