LOCUS       sequence-445204-        6976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6976)
  AUTHORS   Braun JE, Huntzinger E, Fauser M, Izaurralde E
  TITLE     GW182 proteins directly recruit cytoplasmic deadenylase complexes to
            miRNA targets.
  JOURNAL   Mol Cell. 2011 Oct 7;44(1):120-33.
  PUBMED    21981923
REFERENCE   2  (bases 1 to 6976)
  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"; journalName: "Mol Cell.";
            date: "2011-10-7"; volume: "44(1)"; pages: "120-33"
COMMENT     Sequence Label: pT7-EGFP-C1-HsNot10_K
FEATURES             Location/Qualifiers
     source          1..6976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /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(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3704..3825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3741..3760)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3795..3814
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3832..4287)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3969..3990)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4181..4200
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4314..4418
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4420..4777
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4420..4611
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4443..4463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4628..4763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4812..5606
                     /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             4812..5606
                     /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(4866..4885)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5476..5495
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5794..5813)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5838..5885
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6214..6802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6703..6722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag ctgctgcaga caagcctgca gatcagggag cagagaaaca
     1381 tgaaggcaca ggtcagtcct ctgggatcac tgatcaagag aaggagttat ccaccaatgc
     1441 tttccaagct ttcacatctg gaaattatga tgcctgtcta caacaccttg cctgtctaca
     1501 agatataaac aaagatgatt ataaaataat tttgaataca gcagtagctg agttttttaa
     1561 aagtaaccaa acaacaacag ataatttgag acaaacactt aaccagctga agaatcaggt
     1621 ccactcagct gttgaagaaa tggatggatt agatgatgtt gaaaacagca tgttgtacta
     1681 taatcaagca gtcattcttt atcatctgcg gcagtataca gaagccatat cagttggtga
     1741 aaaactttat cagttcatag agccttttga agaaaaattt gcccaagcag tgtgtttttt
     1801 gcttgtagac ctgtatatat taacctacca agctgagaaa gctttgcatc ttcttgctgt
     1861 cctagaaaaa atgatttcac agggtaacaa taacaaaaat ggaaagaatg agactggtaa
     1921 taacaacaac aaagatggat ctaatcataa agctgaaagt ggagctctaa tagaagctgc
     1981 aaaatcaaag atacatcagt acaaagtacg agcttatatc caaatgaagt ctctgaaagc
     2041 atgtaaaagg gaaatcaagt cagtcatgaa tacagctgga aattccgcac cctctctctt
     2101 tcttaaaagc aattttgagt acttaagagg taattatcga aaagccgtga agctattaaa
     2161 tagttcaaac attgctgagc atccaggatt catgaaaaca ggtgaatgct tgagatgcat
     2221 gttctggaat aaccttggtt gcatccattt tgccatgagc aagcacaatt tgggaatatt
     2281 ctactttaaa aaggctctgc aagagaatga caatgtctgt gcacagctca gtgcaggtag
     2341 cactgatcca ggtaaaaaat tttcaggaag acccatgtgt acgttactaa ccaataagag
     2401 atatgagttg ctgtataact gtggaattca gcttcttcac attggaaggc ctcttgctgc
     2461 cttcgaatgt ctgattgaag ctgttcaggt ttatcatgca aatcctcgcc tctggctacg
     2521 gctggctgaa tgctgcattg ctgccaataa ggggacttct gaacaagaaa ctaaaggcct
     2581 tcccagcaaa aaaggaattg tacagtctat tgttggtcaa ggctatcatc gtaaaatagt
     2641 tttggcatca cagtctatac agaatactgt ttataatgat ggtcagtctt cggccattcc
     2701 tgtagccagt atggagtttg cagccatatg tctcagaaat gccttgttgc tgctacctga
     2761 agaacagcaa gatccaaagc aggaaaatgg ggctaaaaat agtaatcaat taggtgggaa
     2821 cacagagagc agcgaaagca gtgaaacttg cagcagtaaa agccatgatg gagataaatt
     2881 cattccagct ccaccttctt ctccattgag aaaacaggaa ttagaaaact taaagtgctc
     2941 catacttgct tgcagtgcct acgtggctct ggctttgggt gataacctca tggctttgaa
     3001 tcatgcagat aaacttcttc agcagcccaa gctgtcagga tctcttaagt ttttgggaca
     3061 tttatatgct gcagaagccc tcatctctct cgacagaata tctgatgcca ttactcactt
     3121 gaacccggag aatgtcactg atgtctcctt agggatctct tcaaatgagc aggaccaagg
     3181 atcagacaaa ggtgaaaatg aagcaatgga atcctctggt aagcgggccc ctcagtgcta
     3241 ccccagttcc gtcaactctg ccaggactgt gatgctgttc aaccttggca gcgcttactg
     3301 cctgaggagc gaatatgaca aagcccgaaa gtgtctccac caggcggctt caatgatcca
     3361 tcctaaagag gtgccccctg aggccatctt gctggcagtc taccttgaac tgcagaatgg
     3421 taatactcag ctggccttac agatcatcaa aaggaatcag ctgctccctg cagtgaaaac
     3481 acactctgaa gtgagaaaga agccagtgtt tcagcctgtc cacccgatcc agcccatcca
     3541 aatgccggct ttcaccactg tgcagagaaa gtgaggatcc accggatcta gataactgat
     3601 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3661 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3721 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3781 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     3841 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3901 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3961 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4021 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4081 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4141 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4201 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4261 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4321 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4381 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4441 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4501 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4561 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4621 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4681 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4741 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4801 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4861 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4921 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4981 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5041 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5101 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5161 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5221 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5281 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5341 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5401 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5461 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5521 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5581 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5641 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5701 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5761 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5821 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5881 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5941 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6001 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6061 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6121 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6841 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6901 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6961 catatatgga gttccg
//
