LOCUS       sequence-290270-        7619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUpf1-T28A-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7619)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7619)
  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: pT7-V5-SBP-C1-HsUpf1-T28A-siRNAres_W
FEATURES             Location/Qualifiers
     source          1..7619
                     /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             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /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="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    4407..4528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4444..4463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4498..4517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4535..4990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4672..4693)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4884..4903
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5017..5121
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5123..5480
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5123..5314
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5146..5166)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5331..5466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5515..6309
                     /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             5515..6309
                     /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(5569..5588)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6179..6198
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6497..6516)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6541..6588
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6917..7505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7406..7425
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctatga gcgtggaggc gtacgggccc agctcgcaga ctctcacttt
      901 cctggacacg gaggaggccg agctgcttgg cgccgacgca cagggctccg agttcgagtt
      961 caccgacttt actcttccta gccagacgca gacgcccccc ggcggccccg gcggcccggg
     1021 cggtggcggc gcgggaggcc cgggcggcgc gggcgcgggc tctgcggcgg gacagctcga
     1081 cgcgcaggtt gggcccgaag gcatcctgca gaacggggct gtggacgaca gtgtagccaa
     1141 gaccagccag ttgttggctg agttgaactt cgaggaagat gaagaagaca cctattacac
     1201 gaaggacctc cccatacacg cctgcagtta ctgtggaata cacgatcctg cctgcgtggt
     1261 ttactgtaat accagcaaga agtggttctg caacggacgt ggaaatactt ctggcagcca
     1321 cattgtaaat caccttgtga gggcaaaatg caaaggggtg accctgcaca aggacgggcc
     1381 cctgggggag acagtcctgg agtgctacaa ctgcggctgt cgcaacgtct tcctcctcgg
     1441 cttcatcccg gccaaagctg actcagtggt ggtgctgctg tgcaggcagc cctgtgccag
     1501 ccagagcagc ctcaaggaca tcaactggga cagctcgcag tggcagccgc tgatccagga
     1561 ccgctgcttc ctgtcctggc tggtcaagat cccctccgag caggagcagc tgcgggcacg
     1621 ccagatcacg gcacagcaga tcaacaagct ggaggagctg tggaaggaaa acccttctgc
     1681 cacgctggag gacctggaga ggccgggggt ggacgaggag ccgcagcatg tcctcctgcg
     1741 gtacgaggac gcctaccagt accagaacat attcgggccc ctggtcaagc tggaggccga
     1801 ctacgacaag aagctgaagg agtcccagac tcaagataac atcactgtca ggtgggacct
     1861 gggccttaac aagaagagaa tcgcctactt cactttgccc aagactgact ctggtaatga
     1921 ggatttagtc ataatttggt taagagacat gcggctcatg cagggggatg agatatgcct
     1981 gcggtacaaa ggggaccttg cgcccctgtg gaaagggatc ggccacgtca tcaaggtccc
     2041 tgataattat ggcgatgaga tcgccattga gctgcggagc agcgtgggtg cacctgtgga
     2101 ggtgactcac aacttccagg tggattttgt gtggaagtcg acctcctttg acaggatgca
     2161 gagcgcattg aaaacgtttg gcgtggatga gacctcggtg tctggctaca tctaccacaa
     2221 gctgttgggc cacgaggtgg aggacgtaat catcaagtgc cagctgccca agcgcttcac
     2281 ggcgcagggc ctccccgacc tcaaccactc ccaggtttat gccgtgaaga ctgtgctgca
     2341 aagaccactg agcctgatcc agggcccgcc aggcacgggg aagacggtga cgtcggccac
     2401 catcgtctac cacctggccc ggcaaggcaa cgggccggtg ctggtgtgtg ctccgagcaa
     2461 catcgccgtg gaccagctaa cggagaagat ccaccagacg gggctaaagg tcgtgcgcct
     2521 ctgcgccaag agccgtgagg ccatcgactc cccggtgtct tttctggccc tgcacaacca
     2581 gatcaggaac atggacagca tgcctgagct gcagaagctg cagcagctga aagacgagac
     2641 tggggagctg tcgtctgccg acgagaagcg gtaccgggcc ttgaagcgca ccgcagagag
     2701 agagctgctg atgaacgcag atgtcatctg ctgcacatgt gtgggcgccg gtgacccgag
     2761 gctggccaaa atgcaattta ggagtatttt aatcgacgaa agcacccagg ccaccgagcc
     2821 ggagtgcatg gttcccgtgg tcctcggggc caagcagctg atccttgtag gcgaccactg
     2881 ccagctgggc ccagtggtga tgtgcaagaa ggcggccaag gccgggctgt cacagtcgct
     2941 cttcgagcgc ctggtggtgc tgggcatccg gcccatccgc ctgcaggtcc agtaccggat
     3001 gcaccctgca ctcagcgcct tcccatccaa catcttctac gagggctccc tccagaatgg
     3061 tgtcactgca gcggatcgtg tgaagaaggg atttgacttc cagtggcccc aacccgataa
     3121 accgatgttc ttctacgtga cccagggcca agaggagatt gccagctcgg gcacctccta
     3181 cctgaacagg accgaggctg cgaacgtgga gaagatcacc acgaagttgc tgaaggcagg
     3241 cgccaagccg gaccagattg gcatcatcac gccctacgag ggccagcgct cctacctggt
     3301 gcagtacatg cagttcagcg gctccctgca caccaagctc taccaggagg tggagatcgc
     3361 cagtgtggac gcctttcagg gacgcgagaa ggacttcatc atcctgtcct gtgtgcgggc
     3421 caacgagcac caaggcattg gctttttaaa tgaccccagg cgtctgaacg tggccctgac
     3481 cagagcaagg tatggcgtca tcattgtggg caacccgaag gcactatcaa agcagccgct
     3541 ctggaaccac ctgctgaact actataagga gcagaaggtg ctggtggagg ggccgctcaa
     3601 caacctgcgt gagagcctca tgcagttcag caagccacgg aagctggtca acactatcaa
     3661 cccgggagcc cgcttcatga ccacagccat gtatgatgcc cgggaggcca tcatcccagg
     3721 ctccgtctat gatcggagca gccagggccg gccttccagc atgtacttcc agacccatga
     3781 ccagattggc atgatcagtg ccggccctag ccacgtggct gccatgaaca ttcccatccc
     3841 cttcaacctg gtcatgccac ccatgccacc gcctggctat tttggacaag ccaacgggcc
     3901 tgctgcaggg cgaggcaccc cgaaaggcaa gactggtcgt gggggacgcc agaagaaccg
     3961 ctttgggctt cctggaccca gccagactaa cctccccaac agccaagcca gccaggatgt
     4021 ggcgtcacag cccttctctc agggcgccct gacgcagggc tacatctcca tgagccagcc
     4081 ttcccagatg agccagcccg gcctctccca gccggagctg tcccaggaca gttaccttgg
     4141 tgacgagttt aaatcacaaa tcgacgtggc gctctcacag gactccacgt accagggaga
     4201 gcgggcttac cagcatggcg gggtgatggg gctgtcccag tattaagaat tctgcagtcg
     4261 acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     4321 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4381 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4501 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     4561 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4621 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4681 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4741 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4801 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4861 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4921 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4981 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5101 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     5161 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5221 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5281 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5341 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5401 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5461 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5521 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5581 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5641 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5701 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5761 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5821 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5881 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5941 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6001 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     6061 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6121 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6181 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6241 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6301 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6361 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6421 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6481 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6541 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6601 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6661 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6721 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7561 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
