LOCUS       sequence-356534-        7988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUPF2-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7988)
  AUTHORS   Kashima I, Jonas S, Jayachandran U, Buchwald G, Conti E, Lupas AN, 
            Izaurralde E
  TITLE     SMG6 interacts with the exon junction complex via two conserved 
            EJC-binding motifs (EBMs) required for nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2010 Nov 1. 24(21):2440-50.
  PUBMED    20930030
REFERENCE   2  (bases 1 to 7988)
  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"; journalName: "Genes Dev. 
            2010 Nov 1. 24(21):2440-50."
COMMENT     Sequence Label: pLNHA-C1-HsUPF2-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..7988
                     /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"
     RBS             1550..1561
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             2323..2331
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4776..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4813..4832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4867..4886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4904..5359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5041..5062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5253..5272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5386..5490
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5492..5849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5492..5683
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5515..5535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5700..5835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5884..6678
                     /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             5884..6678
                     /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(5938..5957)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6548..6567
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6866..6885)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6910..6957
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7286..7874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7775..7794
                     /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 cgagctatgc cagctgagcg taaaaagcca gcaagtatgg aagaaaaaga
      841 ctctttacca aacaacaagg aaaaagactg cagtgaaagg cggacagtga gcagcaagga
      901 gaggccaaaa gacgatatca agctcactgc caagaaggag gtcagcaagg cccctgaaga
      961 caagaagaag agactggaag atgataagag aaaaaaggaa gataaagagc ggaagaaaaa
     1021 agacgaagaa aaggtgaagg cagaggaaga atcaaagaaa aaagaagagg aagaaaaaaa
     1081 gaaacatcaa gaggaagaga gaaagaagca agaagagcag gccaaacgtc agcaagaaga
     1141 agaagcagct gctcagatga aagaaaaaga agaatccatt cagcttcatc aggaagcttg
     1201 ggaacgacat catttaagaa aggaacttcg tagcaaaaac caaaatgctc cggacagccg
     1261 accagaggaa aacttcttca gccgcctcga ctcaagtttg aagaaaaata ctgcttttgt
     1321 caagaaacta aaaactatta cagaacaaca gagagactcc ttgtcccatg attttaatgg
     1381 cctaaattta agcaaataca ttgcagaagc tgtagcttcc atcgtggaag caaaactaaa
     1441 aatctctgat gtgaactgtg ctgtgcacct ctgctctctc tttcaccagc gttatgctga
     1501 ctttgcccca tcacttcttc aggtctggaa aaaacatttt gaagcaagga aagaggagaa
     1561 aacacctaac atcaccaagt taagaactga tttgcgtttt attgcagaat tgacaatagt
     1621 tgggattttc actgacaagg aaggtctttc cttaatctat gaacagctaa aaaatattat
     1681 taatgctgat cgggagtccc acactcatgt ctctgtagtg attagtttct gtcgacattg
     1741 tggagatgat attgctggac ttgtaccaag gaaagtaaag agtgctgcag agaagtttaa
     1801 tttgagtttt cctcctagtg agataattag tccagagaaa caacagccct tccagaatct
     1861 tttaaaagag tactttacgt ctttgaccaa acacctgaaa agggaccaca gggagctcca
     1921 gaatactgag agacaaaaca ggcgcattct acattctaaa ggggagctca gtgaagatag
     1981 acataaacag tatgaggaat ttgctatgtc ttaccagaag ctgctggcaa attctcaatc
     2041 cttagcagac cttttggatg aaaatatgcc agatcttcct caagacaaac caacaccaga
     2101 agaacatggg cctggaattg atatattcac acctggtaaa cctggagaat atgacttgga
     2161 aggtggtata tgggaagatg aagatgctcg gaatttttat gagaacctca ttgatttgaa
     2221 ggcttttgtc ccagccatct tgtttaaaga caatgaaaaa agttgtcaga ataaagagtc
     2281 caacaaagat gataccaaag aggcaaaaga atctaaggag aataaggagg tatcaagtcc
     2341 cgatgatttg gaacttgagt tggagaatct agaaattaat gatgacacct tagaattaga
     2401 gggtggagat gaagctgaag atcttacaaa gaaacttctt gatgaacaag aacaagaaga
     2461 tgaggaagcc agcactggat ctcatctcaa gctcatagta gatgctttcc tacaacagtt
     2521 acccaactgt gtcaaccgag atctgataga caaggcagca atggattttt gcatgaacat
     2581 gaacacaaaa gcaaacagga agaagttggt acgggcactc ttcatagttc ctagacaaag
     2641 gttggatttg ctaccatttt atgcaagatt ggttgctaca ttgcatccct gcatgtctga
     2701 tgtagcagag gatctttgtt ccatgctgag gggggatttc agatttcatg tacggaaaaa
     2761 ggaccagatc aatattgaaa caaagaataa aactgttcgt tttataggag aactaactaa
     2821 gtttaagatg ttcaccaaaa atgacacact gcattgttta aagatgcttc tgtcagactt
     2881 ctctcatcac catattgaaa tggcatgcac cctgctggag acatgtggac ggtttctttt
     2941 cagatctcca gaatctcacc tgaggaccag tgtacttttg gagcaaatga tgagaaagaa
     3001 gcaagcaatg catcttgatg cgagatacgt cacaatggta gagaatgcat attactactg
     3061 caacccacct ccagctgaaa aaaccgtgaa aaagaaacgt cctcctctcc aggaatatgt
     3121 ccggaaactt ttgtacaagg atctctctaa ggttaccacc gagaaggttt tgagacagat
     3181 gcgaaagctg ccctggcagg accaagaagt gaaagactat gttatttgtt gtatgataaa
     3241 catctggaat gtgaaatata atagtattca ttgtgtagcc aacctcttag caggactagt
     3301 gctctaccaa gaggatgttg ggatccacgt tgtggatgga gtgttagaag atattcgatt
     3361 aggaatggag gttaatcaac ctaaatttaa tcagaggcgc atcagcagtg ccaagttctt
     3421 aggagaactt tacaattacc gaatggtgga atcagctgtt attttcagaa ctctgtattc
     3481 ttttacctca tttggtgtta atcctgatgg ctctccaagt tccctggacc cacctgagca
     3541 tcttttcaga attagactcg tatgcactat tctggacaca tgtggccagt actttgacag
     3601 aggttccagt aaacgaaaac ttgattgttt ccttgtatat tttcagcgtt atgtttggtg
     3661 gaagaaaagt ttggaggttt ggacaaaaga ccatccattt cctattgata tagattacat
     3721 gatcagtgat acactagaac tgctaagacc aaagatcaaa ctctgtaatt ctctggaaga
     3781 atccatcagg caggtacaag acttggaacg agaattctta ataaaactag gcctagtaaa
     3841 tgacaaagac tcaaaagatt ctatgacaga aggagaaaat cttgaagagg atgaagaaga
     3901 agaagaaggt ggggctgaaa cagaagaaca atctggaaat gaaagtgaag taaatgagcc
     3961 agaagaagag gagggttctg ataatgatga tgatgaggga gaagaagagg aggaagagaa
     4021 tacagattac cttacagatt ccaataagga aaatgaaacc gatgaagaga atactgaggt
     4081 aatgattaaa ggcggtggac ttaagcatgt accttgtgta gaagatgagg acttcattca
     4141 agctctggat aaaatgatgc tagaaaatct acagcaacga agtggtgagt ctgttaaagt
     4201 gcaccaacta gatgttgcca ttcctttgca tctcaaaagc cagctgagga aagggccccc
     4261 actgggaggt ggggaaggag aggctgagtc tgcagacaca atgccgtttg tcatgttaac
     4321 aagaaaaggc aataaacagc agtttaagat ccttaatgta cccatgtcct ctcaacttgc
     4381 tgcaaatcac tggaaccagc aacaggcaga acaagaagag aggatgagaa tgaaaaagct
     4441 cacactagat atcaatgaac ggcaagaaca agaagattat caagaaatgt tgcagtctct
     4501 tgcacagcgc ccagctccag caaacaccaa tcgtgagagg cggcctcgct accaacatcc
     4561 gaagggagca cctaatgcag atctaatctt taagactggt gggaggagac gttgacaatt
     4621 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     4681 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4741 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5341 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5401 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5461 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     5521 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5581 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5641 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5701 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5761 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5821 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5881 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5941 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6001 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6061 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6121 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6181 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6241 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6301 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6361 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     6421 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6481 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6541 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6601 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6661 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6721 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6781 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6841 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6901 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6961 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     7021 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     7081 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     7141 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7201 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7921 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7981 ccatgcat
//
