LOCUS       sequence-289253-        8007 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1-1089.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8007)
  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 8007)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_1-1089_K
FEATURES             Location/Qualifiers
     source          1..8007
                     /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    4795..4916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4832..4851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4886..4905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4923..5378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5060..5081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5272..5291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5405..5509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5511..5868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5511..5702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5534..5554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5719..5854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5903..6697
                     /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             5903..6697
                     /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(5957..5976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6567..6586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6885..6904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6929..6976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7305..7893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7794..7813
                     /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 ctaatcttga ctcgctctcg ctggccttgt ctcaaatcag
     1441 ctacctggtg gacaatttaa ccaagaaaaa ttaccgagcc agccagcagg aaatacagca
     1501 tattgtgaat cggcacggtc ctgaggcaga caggcattta ttacgctgcc tattttcgca
     1561 tgtggatttc agtggcgatg gtaaaagcag tggcaaagat ttccatcaga ctcagtttct
     1621 gattcaggag tgtgcgttgc tgattacaaa gccaaatttt atctcgacgc tgtcctatgc
     1681 cattgataat ccattgcact atcagaagag tttaaagcct gcaccccact tatttgccca
     1741 gctgagtaaa gtgctcaaat taagcaaagt acaagaggta atttttggcc ttgccctgtt
     1801 gaattcttcc agctcagatc ttagaggttt cgctgcccag tttatcaaac agaagcttcc
     1861 agatcttctg cgttcttaca ttgacgcaga cgtcagtgga aatcaagaag gtggcttcca
     1921 agatatagca atagaggtcc tacacctcct cctctcccat ctcctctttg ggcagaaggg
     1981 agcctttgga gttggacaag aacagataga cgcttttctt aagacgctgc gcagagattt
     2041 tccccaagaa cgctgtcccg tggtgctcgc accactttta taccctgaaa aacgggacat
     2101 tctaatggac aggatcctgc ctgattccgg aggggtagct aaaaccatga tggagagctc
     2161 tttggctgat ttcatgcaag aagtaggcta tggcttttgt gcaagtattg aagaatgtcg
     2221 caatataatc gtgcagtttg gtgttcggga ggtcacagct gcccaggttg caagggtttt
     2281 gggaatgatg gctcgaactc attcaggatt aacagatggc attccattac agagtatttc
     2341 tgctccgggc agtgggatct ggagtgatgg gaaagataaa agtgatggag cacaggcaca
     2401 cacatggaat gtagaagtct tgattgacgt tcttaaagaa ctgaatccaa gtttgaattt
     2461 caaggaagta acttatgaac tggaccatcc tggatttcaa attcgtgaca gtaaaggact
     2521 tcataatgtg gtttatggca ttcagagggg tttgggtatg gaagtgttcc cagtagacct
     2581 catatataga ccttggaaac atgctgaagg ccagctctcc ttcattcaac attcccttat
     2641 aaatccagag atcttctgtt ttgctgacta tccctgtcat actgttgcca ctgatattct
     2701 gaaagcacca ccagaggatg acaatcgaga aattgccaca tggaagagct tggatttgat
     2761 tgaatctctg ctgaggcttg cagaggttgg gcagtatgag caagtcaaac agctcttcag
     2821 cttccctatc aaacactgtc cagacatgct ggtattggcc ttactacaaa ttaacacctc
     2881 ttggcatacc ttgcgccatg aacttatctc cactctgatg ccaattttcc ttggaaacca
     2941 tcctaactca gctattattt tgcactatgc atggcatggg cagggacagt ctccctcaat
     3001 tcgccaactt atcatgcatg caatggcaga atggtacatg agaggggagc agtatgatca
     3061 ggccaaattg tctcgaatac ttgatgtggc ccaggacttg aaggccttgt caatgctgct
     3121 aaatggtact ccatttgcct ttgttattga ccttgctgca cttgcttcac gtcgtgaata
     3181 cctcaaactt gataagtggc tcacagataa aattcgagag catggggagc cttttatcca
     3241 ggcgtgtatg acttttttaa agagacggtg tccttctatt ttgggcggac ttgccccaga
     3301 aaaagaccag cccaaaagtg ctcaacttcc tccagaaact ttggcgacaa tgttggcctg
     3361 tctgcaagct tgtgcaggga gtgtttctca ggagctatca gaaactatcc tcaccatggt
     3421 agccaattgc agtaatgtta tgaataaggc cagacaacca ccacctggag ttatgccaaa
     3481 aggacgtcct cctagtgcta gcagcttaga tgccatttct cctgttcaga ttgaccctct
     3541 tgctggaatg acatctctta gtataggtgg ttcagctgcc cctcacaccc agagtatgca
     3601 gggttttcct ccaaatttgg gttctgcatt cagtacccct cagtcaccag caaaagcatt
     3661 tccacccctt tcaaccccca atcagaccac tgcattcagt ggtattggag gactttcatc
     3721 acagcttcca gtaggtggtc ttggcacagg cagcctgact ggtataggaa ctggtgctct
     3781 tggactccct gcagtgaata acgacccttt tgtacagagg aaactgggca cctctggact
     3841 gaatcagcct acattccagc agacggactt gtctcaggtg tggccagagg caaaccagca
     3901 ctttagtaaa gagatagatg atgaagcaaa cagctatttc cagcgaatat ataatcatcc
     3961 accacatcca accatgtctg ttgatgaggt attagaaatg ctgcagagat ttaaagactc
     4021 tactataaag agggaacgag aagtatttaa ctgtatgcta aggaacttgt ttgaagaata
     4081 tcgttttttt ccccagtatc ctgataaaga gttacatata acagcctgcc tatttggtgg
     4141 tataattgag aaaggactgg tcacttacat ggcactaggt ctggctctac gatatgttct
     4201 tgaagcctta cgcaagcctt ttggatccaa aatgtattat ttcgggattg ctgcactaga
     4261 tagatttaaa aacagattga aggactatcc ccagtattgt cagcatttgg cttctatcag
     4321 tcactttatg caatttccac atcatttaca ggagtatatt gagtatggac agcagtctag
     4381 agatcctcct gtgaaaatgc aaggctctat cacaacccct ggaagtattg cactggctca
     4441 ggcccaggct caggcccagg ttccagcaaa agctcctctt gctggtcaag ttagcactat
     4501 ggtaaccacc tcaacaacta ccactgttgc taaaacggtt acggtcacca ggccaactgg
     4561 agtcagcttt aagaaagatg tgccaccttc tattaatact acaaatatag atacgttgct
     4621 tgtggccaca gatcaaactg agagaattgt gtgaccgcgg gcccgggatc caccggatct
     4681 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4741 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4801 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4861 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     4921 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4981 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5041 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5101 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5161 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5221 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5281 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5341 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     5521 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5821 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     5881 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5941 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     6001 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6061 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     6121 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6181 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6241 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6301 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6361 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6421 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6481 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6541 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6601 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6661 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     6721 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     6781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6841 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     6901 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     6961 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     7021 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     7081 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     7141 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7441 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7981 gtggataacc gtattaccgc catgcat
//
