LOCUS       sequence-289986-        7803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1356-2370.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7803)
  AUTHORS   Bawankar P, Loh B, Wohlbold L, Schmidt S, Izaurralde E
  TITLE     NOT10 and C2orf29/NOT11 form a conserved module of the CCR4-NOT 
            complex that docks onto the NOT1 N-terminal domain.
  JOURNAL   RNA Biol. 2013 Feb;10(2):228-44. doi: 10.4161/rna.23018. Epub 2013 
            Jan 9.
  PUBMED    23303381
REFERENCE   2  (bases 1 to 7803)
  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: "10.4161/rna.23018";
            journalName: "RNA Biol"; date: "2013-02"; volume: "10"; issue: "2"; 
            pages: "228-44"
COMMENT     Sequence Label: pT7-EGFP-C1-HsNot1iso1_1356-2370_S
FEATURES             Location/Qualifiers
     source          1..7803
                     /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    4591..4712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4628..4647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4682..4701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4719..5174)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4856..4877)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5068..5087
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5201..5305
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5307..5664
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5307..5498
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5330..5350)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5515..5650
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5699..6493
                     /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             5699..6493
                     /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(5753..5772)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6363..6382
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6681..6700)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6725..6772
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7101..7689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7590..7609
                     /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 ctcagtacag ctaccacgac atcaatgtct attcccttgc
     1441 gggcttggca ccacacatta ctctaaatcc aacaattccc ttgtttcagg cccatccaca
     1501 gttgaagcag tgtgtgcgtc aggcaattga acgggctgtc caggagctgg tccatcctgt
     1561 ggtggatcga tcaattaaga ttgccatgac tacttgtgag caaatagtca ggaaggattt
     1621 tgccctggat tcggaggaat ctcgaatgcg aatagcagct catcacatga tgcgtaactt
     1681 gacagctgga atggctatga ttacatgcag ggaacctttg ctcatgagca tatctaccaa
     1741 cttaaaaaac agttttgcct cagcccttcg tactgcttcc ccacaacaaa gagaaatgat
     1801 ggatcaggca gctgctcaat tagctcagga caattgtgag ttggcttgct gttttattca
     1861 gaagactgca gtagaaaaag caggccctga gatggacaag agattagcaa ctgaatttga
     1921 gctgagaaaa catgctaggc aagaaggacg cagatactgt gatcctgttg ttttaacata
     1981 tcaagctgaa cggatgccag agcaaatcag gctgaaagtt ggtggtgtgg acccaaagca
     2041 gttggctgtt tacgaagagt ttgcacgcaa tgttcctggc ttcttaccta caaatgactt
     2101 aagtcagccc acgggatttt tagcccagcc catgaagcaa gcttgggcaa cagatgatgt
     2161 agctcagatt tatgataagt gtattacaga actggagcaa catctacatg ccatcccacc
     2221 aactttggcc atgaaccctc aagctcaggc tcttcgaagt ctcttggagg ttgtagtttt
     2281 atctcgaaac tctcgggatg ccatagctgc tcttggattg ctccaaaagg ctgtagaggg
     2341 cttactagat gccacaagtg gtgctgatgc tgaccttctg ctgcgctaca gggaatgcca
     2401 cctcttggtc ctaaaagctc tgcaggatgg ccgggcatat gggtctccat ggtgcaacaa
     2461 acagatcaca aggtgcctaa ttgaatgtcg agatgaatat aaatataatg tggaggctgt
     2521 ggagctgcta attcgcaatc atttggttaa tatgcagcag tatgatcttc acctagcgca
     2581 gtcaatggag aatggcttaa actacatggc tgtggcattt gctatgcagt tagtaaaaat
     2641 cctgctggtg gatgaaagga gtgttgctca tgttactgag gcagatctgt tccacaccat
     2701 tgaaaccctc atgaggatta atgctcattc cagaggcaat gctccagaag gattgcccca
     2761 gctgatggaa gtagtgcgat ccaactatga agcaatgatt gatcgtgctc atggaggccc
     2821 aaactttatg atgcattctg ggatctctca agcctcagag tatgatgacc ctccaggcct
     2881 gagggagaag gcagagtatc ttctgaggga atgggtgaat ctctaccatt cagcagcagc
     2941 tggccgcgac agtaccaaag ctttctctgc atttgttgga cagatgcacc agcaaggaat
     3001 actgaagacc gatgatctca taacaaggtt ctttcgtctg tgtactgaaa tgtgtgttga
     3061 aatcagttac cgtgctcagg ctgagcagca gcacaatcct gctgccaatc ccaccatgat
     3121 ccgagccaag tgctatcaca acctggatgc ctttgttcga ctcattgcac tgctcgtgaa
     3181 acactcaggg gaggccacca acactgtcac aaagattaat ctgctgaaca aggtccttgg
     3241 tatagtagtg ggagttctcc ttcaggatca tgatgttcgt cagagtgaat ttcagcaact
     3301 tccctaccat cgaattttta tcatgcttct cttggaactc aatgcacctg agcatgtgtt
     3361 ggaaaccatt aatttccaga cacttacagc tttctgcaat acattccaca tcttgaggcc
     3421 taccaaagct cctggctttg tatatgcctg gcttgaactg atttcccatc ggatatttat
     3481 tgcaagaatg ctggcacata cgccacagca gaaggggtgg cctatgtatg cacagctact
     3541 gattgattta ttcaaatatt tagcgccttt ccttagaaat gtggaactca ccaaacctat
     3601 gcaaatcctc tacaagggca ctttaagagt gctgctggtt cttttgcatg atttcccaga
     3661 gttcctttgt gattaccatt atgggttctg tgatgtgatc ccacctaatt gtatccagtt
     3721 aagaaatttg atcctgagtg cctttccaag aaacatgagg ctccccgacc cattcactcc
     3781 taatctaaag gtggacatgt tgagtgaaat taacattgct ccccggattc tcaccaattt
     3841 cactggagta atgccacctc agttcaaaaa ggatttggat tcctatctta aaactcgatc
     3901 accagtcact ttcctgtctg atctgcgcag caacctacag gtatccaatg aacctgggaa
     3961 tcgctacaac ctccagctca tcaatgcact ggtgctctat gtcgggactc aggccattgc
     4021 gcacatccac aacaagggca gcacaccttc aatgagcacc atcactcact cagcacacat
     4081 ggatatcttc cagaatttgg ctgtggactt ggacactgag ggtcgctatc tctttttgaa
     4141 tgcaattgca aatcagctcc ggtacccaaa tagccacact cactacttca gttgcaccat
     4201 gctgtacctt tttgcagagg ccaatacgga agccatccaa gaacagatca caagagttct
     4261 cttggaacgg ttgattgtaa ataggccaca tccttggggt cttcttatta ccttcattga
     4321 gctgattaaa aacccagcgt ttaagttctg gaaccatgaa tttgtacact gtgccccaga
     4381 aatcgaaaag ttattccagt cggtcgcaca gtgctgcatg ggacagaagc aggcccagca
     4441 agtaatgtaa ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4501 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4561 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4621 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4681 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4741 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4801 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4861 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4921 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4981 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5041 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5101 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5161 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5221 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5281 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5341 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5401 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5461 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5521 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5581 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5641 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5701 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5761 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5821 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5881 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5941 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     6001 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     6061 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6121 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6181 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6241 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6301 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6361 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6421 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6481 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6541 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6601 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6661 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6721 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6781 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6841 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6901 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7741 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7801 cat
//
