LOCUS       sequence-430439-        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        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             608..649
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             671..784
                     /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    3885..4006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3922..3941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3976..3995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4013..4468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4150..4171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4362..4381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4601..4958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4601..4792
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4624..4644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4809..4944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4993..5787
                     /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             4993..5787
                     /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(5047..5066)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5657..5676
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5975..5994)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6019..6066
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6395..6983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6884..6903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      661 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      721 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      781 gcccagatct cgagctatga gcgtggaggc gtacgggccc agctcgcaga ctctcacttt
      841 cctggacacg gaggaggccg agctgcttgg cgccgacaca cagggctccg agttcgagtt
      901 caccgacttt actcttccta gccagacgca gacgcccccc ggcggccccg gcggcccggg
      961 cggtggcggc gcgggaggcc cgggcggcgc gggcgcgggc tctgcggcgg gacagctcga
     1021 cgcgcaggtt gggcccgaag gcatcctgca gaacggggct gtggacgaca gtgtagccaa
     1081 gaccagccag ttgttggctg agttgaactt cgaggaagat gaagaagaca cctattacac
     1141 gaaggacctc ccttctgcca cgctggagga cctggagagg ccgggggtgg acgaggagcc
     1201 gcagcatgtc ctcctgcggt acgaggacgc ctaccagtac cagaacatat tcgggcccct
     1261 ggtcaagctg gaggccgact acgacaagaa gctgaaggag tcccagactc aagataacat
     1321 cactgtcagg tgggacctgg gccttaacaa gaagagaatc gcctacttca ctttgcccaa
     1381 gactgactct ggtaatgagg atttagtcat aatttggtta agagacatgc ggctcatgca
     1441 gggggatgag atatgcctgc ggtacaaagg ggaccttgcg cccctgtgga aagggatcgg
     1501 ccacgtcatc aaggtccctg ataattatgg cgatgagatc gccattgagc tgcggagcag
     1561 cgtgggtgca cctgtggagg tgactcacaa cttccaggtg gattttgtgt ggaagtcgac
     1621 ctcctttgac aggatgcaga gcgcattgaa aacgtttggc gtggatgaga cctcggtgtc
     1681 tggctacatc taccacaagc tgttgggcca cgaggtggag gacgtaatca tcaagtgcca
     1741 gctgcccaag cgcttcacgg cgcagggcct ccccgacctc aaccactccc aggtttatgc
     1801 cgtgaagact gtgctgcaaa gaccactgag cctgatccag ggcccgccag gcacggggaa
     1861 gacggtgacg tcggccacca tcgtctacca cctggcccgg caaggcaacg ggccggtgct
     1921 ggtgtgtgct ccgagcaaca tcgccgtgga ccagctaacg gagaagatcc accagacggg
     1981 gctaaaggtc gtgcgcctct gcgccaagag ccgtgaggcc atcgactccc cggtgtcttt
     2041 tctggccctg cacaaccaga tcaggaacat ggacagcatg cctgagctgc agaagctgca
     2101 gcagctgaaa gacgagactg gggagctgtc gtctgccgac gagaagcggt accgggcctt
     2161 gaagcgcacc gcagagagag agctgctgat gaacgcagat gtcatctgct gcacatgtgt
     2221 gggcgccggt gacccgaggc tggccaaaat gcaatttagg agtattttaa tcgacgaaag
     2281 cacccaggcc accgagccgg agtgcatggt tcccgtggtc ctcggggcca agcagctgat
     2341 ccttgtaggc gaccactgcc agctgggccc agtggtgatg tgcaagaagg cggccaaggc
     2401 cgggctgtca cagtcgctct tcgagcgcct ggtggtgctg ggcatccggc ccatccgcct
     2461 gcaggtccag taccggatgc accctgcact cagcgccttc ccatccaaca tcttctacga
     2521 gggctccctc cagaatggtg tcactgcagc ggatcgtgtg aagaagggat ttgacttcca
     2581 gtggccccaa cccgataaac cgatgttctt ctacgtgacc cagggccaag aggagattgc
     2641 cagctcgggc acctcctacc tgaacaggac cgaggctgcg aacgtggaga agatcaccac
     2701 gaagttgctg aaggcaggcg ccaagccgga ccagattggc atcatcacgc cctacgaggg
     2761 ccagcgctcc tacctggtgc agtacatgca gttcagcggc tccctgcaca ccaagctcta
     2821 ccaggaggtg gagatcgcca gtgtggacgc ctttcaggga cgcgagaagg acttcatcat
     2881 cctgtcctgt gtgcgggcca acgagcacca aggcattggc tttttaaatg accccaggcg
     2941 tctgaacgtg gccctgacca gagcaaggta tggcgtcatc attgtgggca acccgaaggc
     3001 actatcaaag cagccgctct ggaaccacct gctgaactac tataaggagc agaaggtgct
     3061 ggtggagggg ccgctcaaca acctgcgtga gagcctcatg cagttcagca agccacggaa
     3121 gctggtcaac actatcaacc cgggagcccg cttcatgacc acagccatgt atgatgcccg
     3181 ggaggccatc atcccaggct ccgtctatga tcggagcagc cagggccggc cttccagcat
     3241 gtacttccag acccatgacc agattggcat gatcagtgcc ggccctagcc acgtggctgc
     3301 catgaacatt cccatcccct tcaacctggt catgccaccc atgccaccgc ctggctattt
     3361 tggacaagcc aacgggcctg ctgcagggcg aggcaccccg aaaggcaaga ctggtcgtgg
     3421 gggacgccag aagaaccgct ttgggcttcc tggacccagc cagactaacc tccccaacag
     3481 ccaagccagc caggatgtgg cgtcacagcc cttctctcag ggcgccctga cgcagggcta
     3541 catctccatg agccagcctt cccagatgag ccagcccggc ctctcccagc cggagctgtc
     3601 ccaggacagt taccttggtg acgagtttaa atcacaaatc gacgtggcgc tctcacagga
     3661 ctccacgtac cagggagagc gggcttacca gcatggcggg gtgatggggc tgtcccagta
     3721 ttaagaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     3781 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3841 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     4021 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4081 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4141 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4201 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4261 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4321 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4381 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4441 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4561 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4621 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4681 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4801 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4861 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4921 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5161 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5521 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5941 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6061 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6121 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6181 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6541 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7081 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     7141 ccatatatgg agttccg
//
