LOCUS       sequence-289894-        6190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot10_25-707.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6190)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6190)
  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: pLNHA-C1-HsNot10_25-707_S
FEATURES             Location/Qualifiers
     source          1..6190
                     /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             665..730
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2978..3099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3015..3034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3069..3088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3106..3561)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3243..3264)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3455..3474
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3588..3692
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3694..4051
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3694..3885
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3717..3737)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3902..4037
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4086..4880
                     /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             4086..4880
                     /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(4140..4159)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4750..4769
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5068..5087)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5112..5159
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5488..6076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5977..5996
                     /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 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctgatc aagagaagga gttatccacc aatgctttcc aagctttcac
      841 atctggaaat tatgatgcct gtctacaaca ccttgcctgt ctacaagata taaacaaaga
      901 tgattataaa ataattttga atacagcagt agctgagttt tttaaaagta accaaacaac
      961 aacagataat ttgagacaaa cacttaacca gctgaagaat caggtccact cagctgttga
     1021 agaaatggat ggattagatg atgttgaaaa cagcatgttg tactataatc aagcagtcat
     1081 tctttatcat ctgcggcagt atacagaagc catatcagtt ggtgaaaaac tttatcagtt
     1141 catagagcct tttgaagaaa aatttgccca agcagtgtgt tttttgcttg tagacctgta
     1201 tatattaacc taccaagctg agaaagcttt gcatcttctt gctgtcctag aaaaaatgat
     1261 ttcacagggt aacaataaca aaaatggaaa gaatgagact ggtaataaca acaacaaaga
     1321 tggatctaat cataaagctg aaagtggagc tctaatagaa gctgcaaaat caaagataca
     1381 tcagtacaaa gtacgagctt atatccaaat gaagtctctg aaagcatgta aaagggaaat
     1441 caagtcagtc atgaatacag ctggaaattc cgcaccctct ctctttctta aaagcaattt
     1501 tgagtactta agaggtaatt atcgaaaagc cgtgaagcta ttaaatagtt caaacattgc
     1561 tgagcatcca ggattcatga aaacaggtga atgcttgaga tgcatgttct ggaataacct
     1621 tggttgcatc cattttgcca tgagcaagca caatttggga atattctact ttaaaaaggc
     1681 tctgcaagag aatgacaatg tctgtgcaca gctcagtgca ggtagcactg atccaggtaa
     1741 aaaattttca ggaagaccca tgtgtacgtt actaaccaat aagagatatg agttgctgta
     1801 taactgtgga attcagcttc ttcacattgg aaggcctctt gctgccttcg aatgtctgat
     1861 tgaagctgtt caggtttatc atgcaaatcc tcgcctctgg ctacggctgg ctgaatgctg
     1921 cattgctgcc aataagggga cttctgaaca agaaactaaa ggccttccca gcaaaaaagg
     1981 aattgtacag tctattgttg gtcaaggcta tcatcgtaaa atagttttgg catcacagtc
     2041 tatacagaat actgtttata atgatggtca gtcttcggcc attcctgtag ccagtatgga
     2101 gtttgcagcc atatgtctca gaaatgcctt gttgctgcta cctgaagaac agcaagatcc
     2161 aaagcaggaa aatggggcta aaaatagtaa tcaattaggt gggaacacag agagcagcga
     2221 aagcagtgaa acttgcagca gtaaaagcca tgatggagat aaattcattc cagctccacc
     2281 ttcttctcca ttgagaaaac aggaattaga aaacttaaag tgctccatac ttgcttgcag
     2341 tgcctacgtg gctctggctt tgggtgataa cctcatggct ttgaatcatg cagataaact
     2401 tcttcagcag cccaagctgt caggatctct taagtttttg ggacatttat atgctgcaga
     2461 agccctcatc tctctcgaca gaatatctga tgccattact cacttgaacc cggagaatgt
     2521 cactgatgtc tccttaggga tctcttcaaa tgagcaggac caaggatcag acaaaggtga
     2581 aaatgaagca atggaatcct ctggtaagcg ggcccctcag tgctacccca gttccgtcaa
     2641 ctctgccagg actgtgatgc tgttcaacct tggcagcgct tactgcctga ggagcgaata
     2701 tgacaaagcc cgaaagtgtc tccaccaggc ggcttcaatg atccatccta aagaggtgcc
     2761 ccctgaggcc atcttgctgg cagtctacct tgaactgcag aatggtaata ctcagctggc
     2821 cttacagatc atcaaaagga atcagtaagg atccaccgga tctagataac tgatcataat
     2881 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2941 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3001 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3061 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     3121 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3181 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3241 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3301 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3361 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3421 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3481 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3541 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3661 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3721 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3781 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3841 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3901 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3961 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4021 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4081 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4141 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4201 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4261 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4321 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4381 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4441 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4501 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4561 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4621 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4681 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4741 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4801 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4861 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4921 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4981 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5041 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5101 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5161 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5221 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5281 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5341 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5401 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5461 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5521 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5581 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5641 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5701 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5761 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5821 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5881 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5941 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6001 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6061 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6121 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6181 cgccatgcat
//
