LOCUS       sequence-289641-        7157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUpf1-del119-272-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7157)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7157)
  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-del119-272-siRNAres_P
FEATURES             Location/Qualifiers
     source          1..7157
                     /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    3945..4066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3982..4001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4036..4055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4073..4528)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4210..4231)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4422..4441
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4555..4659
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4661..5018
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4661..4852
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4684..4704)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4869..5004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5053..5847
                     /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             5053..5847
                     /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(5107..5126)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5717..5736
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6035..6054)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6079..6126
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6455..7043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6944..6963
                     /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 cgccgacaca 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 ccttctgcca cgctggagga cctggagagg ccgggggtgg acgaggagcc
     1261 gcagcatgtc ctcctgcggt acgaggacgc ctaccagtac cagaacatat tcgggcccct
     1321 ggtcaagctg gaggccgact acgacaagaa gctgaaggag tcccagactc aagataacat
     1381 cactgtcagg tgggacctgg gccttaacaa gaagagaatc gcctacttca ctttgcccaa
     1441 gactgactct ggtaatgagg atttagtcat aatttggtta agagacatgc ggctcatgca
     1501 gggggatgag atatgcctgc ggtacaaagg ggaccttgcg cccctgtgga aagggatcgg
     1561 ccacgtcatc aaggtccctg ataattatgg cgatgagatc gccattgagc tgcggagcag
     1621 cgtgggtgca cctgtggagg tgactcacaa cttccaggtg gattttgtgt ggaagtcgac
     1681 ctcctttgac aggatgcaga gcgcattgaa aacgtttggc gtggatgaga cctcggtgtc
     1741 tggctacatc taccacaagc tgttgggcca cgaggtggag gacgtaatca tcaagtgcca
     1801 gctgcccaag cgcttcacgg cgcagggcct ccccgacctc aaccactccc aggtttatgc
     1861 cgtgaagact gtgctgcaaa gaccactgag cctgatccag ggcccgccag gcacggggaa
     1921 gacggtgacg tcggccacca tcgtctacca cctggcccgg caaggcaacg ggccggtgct
     1981 ggtgtgtgct ccgagcaaca tcgccgtgga ccagctaacg gagaagatcc accagacggg
     2041 gctaaaggtc gtgcgcctct gcgccaagag ccgtgaggcc atcgactccc cggtgtcttt
     2101 tctggccctg cacaaccaga tcaggaacat ggacagcatg cctgagctgc agaagctgca
     2161 gcagctgaaa gacgagactg gggagctgtc gtctgccgac gagaagcggt accgggcctt
     2221 gaagcgcacc gcagagagag agctgctgat gaacgcagat gtcatctgct gcacatgtgt
     2281 gggcgccggt gacccgaggc tggccaaaat gcaatttagg agtattttaa tcgacgaaag
     2341 cacccaggcc accgagccgg agtgcatggt tcccgtggtc ctcggggcca agcagctgat
     2401 ccttgtaggc gaccactgcc agctgggccc agtggtgatg tgcaagaagg cggccaaggc
     2461 cgggctgtca cagtcgctct tcgagcgcct ggtggtgctg ggcatccggc ccatccgcct
     2521 gcaggtccag taccggatgc accctgcact cagcgccttc ccatccaaca tcttctacga
     2581 gggctccctc cagaatggtg tcactgcagc ggatcgtgtg aagaagggat ttgacttcca
     2641 gtggccccaa cccgataaac cgatgttctt ctacgtgacc cagggccaag aggagattgc
     2701 cagctcgggc acctcctacc tgaacaggac cgaggctgcg aacgtggaga agatcaccac
     2761 gaagttgctg aaggcaggcg ccaagccgga ccagattggc atcatcacgc cctacgaggg
     2821 ccagcgctcc tacctggtgc agtacatgca gttcagcggc tccctgcaca ccaagctcta
     2881 ccaggaggtg gagatcgcca gtgtggacgc ctttcaggga cgcgagaagg acttcatcat
     2941 cctgtcctgt gtgcgggcca acgagcacca aggcattggc tttttaaatg accccaggcg
     3001 tctgaacgtg gccctgacca gagcaaggta tggcgtcatc attgtgggca acccgaaggc
     3061 actatcaaag cagccgctct ggaaccacct gctgaactac tataaggagc agaaggtgct
     3121 ggtggagggg ccgctcaaca acctgcgtga gagcctcatg cagttcagca agccacggaa
     3181 gctggtcaac actatcaacc cgggagcccg cttcatgacc acagccatgt atgatgcccg
     3241 ggaggccatc atcccaggct ccgtctatga tcggagcagc cagggccggc cttccagcat
     3301 gtacttccag acccatgacc agattggcat gatcagtgcc ggccctagcc acgtggctgc
     3361 catgaacatt cccatcccct tcaacctggt catgccaccc atgccaccgc ctggctattt
     3421 tggacaagcc aacgggcctg ctgcagggcg aggcaccccg aaaggcaaga ctggtcgtgg
     3481 gggacgccag aagaaccgct ttgggcttcc tggacccagc cagactaacc tccccaacag
     3541 ccaagccagc caggatgtgg cgtcacagcc cttctctcag ggcgccctga cgcagggcta
     3601 catctccatg agccagcctt cccagatgag ccagcccggc ctctcccagc cggagctgtc
     3661 ccaggacagt taccttggtg acgagtttaa atcacaaatc gacgtggcgc tctcacagga
     3721 ctccacgtac cagggagagc gggcttacca gcatggcggg gtgatggggc tgtcccagta
     3781 ttaagaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     3841 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3901 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     4081 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4141 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4201 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4261 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4321 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4381 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4441 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4501 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4621 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4681 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4861 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4921 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4981 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5041 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5101 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5161 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5221 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5281 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5341 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5401 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5461 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5521 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5581 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5641 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5701 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5761 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5821 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5881 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5941 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6001 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6061 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6121 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6181 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6241 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6601 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7081 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7141 gtattaccgc catgcat
//
