LOCUS       sequence-291293-        6089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6089)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6089)
  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-HsNot11_AH
FEATURES             Location/Qualifiers
     source          1..6089
                     /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"
     CDS             2195..2305
                     /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             2312..2335
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2369..2542
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2543..2716
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     polyA_signal    2877..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2914..2933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2968..2987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3005..3460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3142..3163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3354..3373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3487..3591
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3593..3950
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3593..3784
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3616..3636)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3801..3936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3985..4779
                     /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             3985..4779
                     /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(4039..4058)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4649..4668
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4967..4986)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5011..5058
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5387..5975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5876..5895
                     /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 tctcgagctc aagcttcgaa ttctgcagtc gacggtacca cagccaccat
      661 gcccggcgga ggggcgagcg cggcgtctgg ccggcttctc accgccgcgg agcaaagagg
      721 gtcccgggaa gcggcagggt cggcgtccag gagcggcttc gggggctccg gcggcggcag
      781 aggcggagca agcggccccg ggtccgggag cggaggcccg gggggccccg cgggcaggat
      841 gagcttgacc ccgaaggagc tctcgagcct gctgagcatc atatcggagg aggcgggcgg
      901 cggcagcacc ttcgagggcc tgtccaccgc cttccaccac tacttcagca aggccgacca
      961 cttccgcctg ggctcggtgc tcgtcatgct gctccagcag cccgacctgc tgcctagcgc
     1021 ggcgcagcgc ctcacggcgc tctacctgct ctgggagatg taccgcaccg agccgctggc
     1081 cgccaacccc ttcgccgcca gcttcgcgca cctgctcaac cccgcgccgc ccgcccgcgg
     1141 cggccaggaa cccgaccgcc ctccgctctc aggattttta cctcctataa ctccaccaga
     1201 aaagtttttt ctttcccagc tgatgctggc acccccacgg gaactcttca aaaagacgcc
     1261 tcgccagatt gcactgatgg acgttggaaa catgggccag tctgtggaca ttagtgggct
     1321 tcagttagcc ttggccgaac gccaatctga attgccaacg caaagcaaag cgagcttccc
     1381 cagtattctc agtgacccag acccggattc ttctaattct ggatttgaca gctcagttgc
     1441 ctctcagatc acagaagctt tagtcagcgg accaaagcca cctattgaaa gccattttcg
     1501 accagagttt attcgtccac cgcctccact ccacatttgt gaggatgaac ttgcttggct
     1561 aaaccccacg gagcctgacc acgcgatcca gtgggataaa tcgatgtgtg ttaagaatag
     1621 cactggtgtg gagatcaaac gaataatggc caaagccttc aaaagcccct tatcctctcc
     1681 ccaacaaaca cagctacttg gtgagttgga aaaagacccc aaacttgtct accatattgg
     1741 cctcacccca gccaaacttc ctgaccttgt ggaaaacaac cctttagtcg ctatagaaat
     1801 gttgctgaaa ttaatgcagt caagccagat cactgagtat ttctctgtcc tggtcaatat
     1861 ggacatgtct ttacattcaa tggaagttgt aaatcgacta actacagctg ttgatctacc
     1921 tcctgaattt attcaccttt atatatcaaa ttgcatctct acttgtgaac agattaagga
     1981 taaatatatg cagaatcggt tggtgcgtct tgtgtgtgtg tttctccaat ccttgatccg
     2041 taacaaaatt attaatgtac aggatttgtt tatagaagtg caggcattct gtattgaatt
     2101 cagtaggata cgagaagctg ctggtctttt ccggttgttg aagacattgg atactgggga
     2161 aacaccttct gagaccaaaa tgtcaaaagg atccgacgag aagaccaccg gctggcgggg
     2221 cggccacgtg gtggagggcc tggccggcga gctggagcag ctgcgggcca ggctggagca
     2281 ccaccctcag ggccagcggg agcccgaact actggaagtt ctgttccagg ggcccaaaac
     2341 cgcggctctt gcgcaacacg atgaagccgt ggacaacaaa ttcaacaaag aacaacaaaa
     2401 cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccttcat
     2461 ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct
     2521 aaatgatgct caggcgccga aagtagacaa caaattcaac aaagaacaac aaaacgcgtt
     2581 ctatgagatc ttacatttac ctaacttaaa cgaagaacaa cgaaacgcct tcatccaaag
     2641 tttaaaagat gacccaagcc aaagcgctaa ccttttagca gaagctaaaa agctaaatga
     2701 tgctcaggcg ccgaaagtag acgcgaattc cgcggggaag tcaaccaagc tttaggcggc
     2761 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2821 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2881 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2941 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     3001 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3061 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3121 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3181 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3241 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3301 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3361 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3421 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3481 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3541 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3601 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3661 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3721 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3781 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3841 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3901 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3961 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4021 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4081 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4141 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4201 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4261 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4321 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4381 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4441 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4501 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4561 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4621 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4681 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4741 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4801 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4861 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4921 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4981 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5041 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5101 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5161 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5221 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5281 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5521 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6061 ctgtggataa ccgtattacc gccatgcat
//
