LOCUS       sequence-289893-        5482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot10_25-471.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5482)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5482)
  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_25-471_S
FEATURES             Location/Qualifiers
     source          1..5482
                     /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    2270..2391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2307..2326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2361..2380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2398..2853)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2535..2556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2747..2766
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2880..2984
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2986..3343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2986..3177
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3009..3029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3194..3329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3378..4172
                     /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             3378..4172
                     /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(3432..3451)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4042..4061
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4360..4379)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4404..4451
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4780..5368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5269..5288
                     /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 gttgctgtaa ggatccaccg gatctagata
     2161 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2221 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2281 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2341 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     2401 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2461 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2521 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2581 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2641 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2701 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2761 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2821 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     2881 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2941 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3001 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3061 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     3361 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3421 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3481 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3541 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3601 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3661 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3721 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3781 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3841 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3901 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3961 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4021 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4081 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4141 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4201 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4261 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4321 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     4381 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     4441 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4501 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4561 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     4621 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4681 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4741 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4801 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4861 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4921 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4981 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5041 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5101 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5161 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5221 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5281 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5341 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5401 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5461 taaccgtatt accgccatgc at
//
