LOCUS       sequence-291292-        6774 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 6774)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6774)
  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"
COMMENT     Sequence Label: pCMV-TAPtagC-PrP-SBP-HsNot10_AH
FEATURES             Location/Qualifiers
     source          1..6774
                     /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"
     regulatory      621..630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2880..2990
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             2997..3020
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3054..3227
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             3228..3401
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     polyA_signal    3562..3683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3599..3618)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3653..3672
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3690..4145)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3848)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4039..4058
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4172..4276
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4278..4635
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4278..4469
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4301..4321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4486..4621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4670..5464
                     /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             4670..5464
                     /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(4724..4743)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5334..5353
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5652..5671)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5696..5743
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6072..6660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6561..6580
                     /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 gcagagctgg tttagtgacc gtcagatccg ctagcgctac
      601 cggactcaga tctcgagaca gccaccatgg ctgcagacaa gcctgcagat cagggagcag
      661 agaaacatga aggcacaggt cagtcctctg ggatcactga tcaagagaag gagttatcca
      721 ccaatgcttt ccaagctttc acatctggaa attatgatgc ctgtctacaa caccttgcct
      781 gtctacaaga tataaacaaa gatgattata aaataatttt gaatacagca gtagctgagt
      841 tttttaaaag taaccaaaca acaacagata atttgagaca aacacttaac cagctgaaga
      901 atcaggtcca ctcagctgtt gaagaaatgg atggattaga tgatgttgaa aacagcatgt
      961 tgtactataa tcaagcagtc attctttatc atctgcggca gtatacagaa gccatatcag
     1021 ttggtgaaaa actttatcag ttcatagagc cttttgaaga aaaatttgcc caagcagtgt
     1081 gttttttgct tgtagacctg tatatattaa cctaccaagc tgagaaagct ttgcatcttc
     1141 ttgctgtcct agaaaaaatg atttcacagg gtaacaataa caaaaatgga aagaatgaga
     1201 ctggtaataa caacaacaaa gatggatcta atcataaagc tgaaagtgga gctctaatag
     1261 aagctgcaaa atcaaagata catcagtaca aagtacgagc ttatatccaa atgaagtctc
     1321 tgaaagcatg taaaagggaa atcaagtcag tcatgaatac agctggaaat tccgcaccct
     1381 ctctctttct taaaagcaat tttgagtact taagaggtaa ttatcgaaaa gccgtgaagc
     1441 tattaaatag ttcaaacatt gctgagcatc caggattcat gaaaacaggt gaatgcttga
     1501 gatgcatgtt ctggaataac cttggttgca tccattttgc catgagcaag cacaatttgg
     1561 gaatattcta ctttaaaaag gctctgcaag agaatgacaa tgtctgtgca cagctcagtg
     1621 caggtagcac tgatccaggt aaaaaatttt caggaagacc catgtgtacg ttactaacca
     1681 ataagagata tgagttgctg tataactgtg gaattcagct tcttcacatt ggaaggcctc
     1741 ttgctgcctt cgaatgtctg attgaagctg ttcaggttta tcatgcaaat cctcgcctct
     1801 ggctacggct ggctgaatgc tgcattgctg ccaataaggg gacttctgaa caagaaacta
     1861 aaggccttcc cagcaaaaaa ggaattgtac agtctattgt tggtcaaggc tatcatcgta
     1921 aaatagtttt ggcatcacag tctatacaga atactgttta taatgatggt cagtcttcgg
     1981 ccattcctgt agccagtatg gagtttgcag ccatatgtct cagaaatgcc ttgttgctgc
     2041 tacctgaaga acagcaagat ccaaagcagg aaaatggggc taaaaatagt aatcaattag
     2101 gtgggaacac agagagcagc gaaagcagtg aaacttgcag cagtaaaagc catgatggag
     2161 ataaattcat tccagctcca ccttcttctc cattgagaaa acaggaatta gaaaacttaa
     2221 agtgctccat acttgcttgc agtgcctacg tggctctggc tttgggtgat aacctcatgg
     2281 ctttgaatca tgcagataaa cttcttcagc agcccaagct gtcaggatct cttaagtttt
     2341 tgggacattt atatgctgca gaagccctca tctctctcga cagaatatct gatgccatta
     2401 ctcacttgaa cccggagaat gtcactgatg tctccttagg gatctcttca aatgagcagg
     2461 accaaggatc agacaaaggt gaaaatgaag caatggaatc ctctggtaag cgggcccctc
     2521 agtgctaccc cagttccgtc aactctgcca ggactgtgat gctgttcaac cttggcagcg
     2581 cttactgcct gaggagcgaa tatgacaaag cccgaaagtg tctccaccag gcggcttcaa
     2641 tgatccatcc taaagaggtg ccccctgagg ccatcttgct ggcagtctac cttgaactgc
     2701 agaatggtaa tactcagctg gccttacaga tcatcaaaag gaatcagctg ctccctgcag
     2761 tgaaaacaca ctctgaagtg agaaagaagc cagtgtttca gcctgtccac ccgatccagc
     2821 ccatccaaat gccggctttc accactgtgc agagaaaggg taccgcgggc ccgggatccg
     2881 acgagaagac caccggctgg cggggcggcc acgtggtgga gggcctggcc ggcgagctgg
     2941 agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc gaactactgg
     3001 aagttctgtt ccaggggccc aaaaccgcgg ctcttgcgca acacgatgaa gccgtggaca
     3061 acaaattcaa caaagaacaa caaaacgcgt tctatgagat cttacattta cctaacttaa
     3121 acgaagaaca acgaaacgcc ttcatccaaa gtttaaaaga tgacccaagc caaagcgcta
     3181 accttttagc agaagctaaa aagctaaatg atgctcaggc gccgaaagta gacaacaaat
     3241 tcaacaaaga acaacaaaac gcgttctatg agatcttaca tttacctaac ttaaacgaag
     3301 aacaacgaaa cgccttcatc caaagtttaa aagatgaccc aagccaaagc gctaaccttt
     3361 tagcagaagc taaaaagcta aatgatgctc aggcgccgaa agtagacgcg aattccgcgg
     3421 ggaagtcaac caagctttag gcggccgcga ctctagatca taatcagcca taccacattt
     3481 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3541 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3601 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3661 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3721 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3781 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3841 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3901 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3961 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4021 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4081 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4141 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4201 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4261 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4381 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4441 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4501 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4561 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4621 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4681 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4741 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4801 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4861 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4921 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4981 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5041 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5101 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5161 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5221 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5281 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5341 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5401 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5461 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5521 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5581 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5641 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5701 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5761 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5821 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5881 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5941 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6001 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6061 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     6241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
