LOCUS       sequence-430435-        7007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUpf1_1-925-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7007)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7007)
  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_1-925-siRNAres_P
FEATURES             Location/Qualifiers
     source          1..7007
                     /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    3735..3856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3772..3791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3826..3845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3863..4318)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4000..4021)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4212..4231
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4345..4449
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4451..4808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4451..4642
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4474..4494)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4659..4794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4843..5637
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGHP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4897..4916)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5507..5526
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5825..5844)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5869..5916
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6245..6833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6734..6753
                     /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 cccatacacg cctgcagtta ctgtggaata cacgatcctg cctgcgtggt
     1201 ttactgtaat accagcaaga agtggttctg caacggacgt ggaaatactt ctggcagcca
     1261 cattgtaaat caccttgtga gggcaaaatg caaaggggtg accctgcaca aggacgggcc
     1321 cctgggggag acagtcctgg agtgctacaa ctgcggctgt cgcaacgtct tcctcctcgg
     1381 cttcatcccg gccaaagctg actcagtggt ggtgctgctg tgcaggcagc cctgtgccag
     1441 ccagagcagc ctcaaggaca tcaactggga cagctcgcag tggcagccgc tgatccagga
     1501 ccgctgcttc ctgtcctggc tggtcaagat cccctccgag caggagcagc tgcgggcacg
     1561 ccagatcacg gcacagcaga tcaacaagct ggaggagctg tggaaggaaa acccttctgc
     1621 cacgctggag gacctggaga ggccgggggt ggacgaggag ccgcagcatg tcctcctgcg
     1681 gtacgaggac gcctaccagt accagaacat attcgggccc ctggtcaagc tggaggccga
     1741 ctacgacaag aagctgaagg agtcccagac tcaagataac atcactgtca ggtgggacct
     1801 gggccttaac aagaagagaa tcgcctactt cactttgccc aagactgact ctggtaatga
     1861 ggatttagtc ataatttggt taagagacat gcggctcatg cagggggatg agatatgcct
     1921 gcggtacaaa ggggaccttg cgcccctgtg gaaagggatc ggccacgtca tcaaggtccc
     1981 tgataattat ggcgatgaga tcgccattga gctgcggagc agcgtgggtg cacctgtgga
     2041 ggtgactcac aacttccagg tggattttgt gtggaagtcg acctcctttg acaggatgca
     2101 gagcgcattg aaaacgtttg gcgtggatga gacctcggtg tctggctaca tctaccacaa
     2161 gctgttgggc cacgaggtgg aggacgtaat catcaagtgc cagctgccca agcgcttcac
     2221 ggcgcagggc ctccccgacc tcaaccactc ccaggtttat gccgtgaaga ctgtgctgca
     2281 aagaccactg agcctgatcc agggcccgcc aggcacgggg aagacggtga cgtcggccac
     2341 catcgtctac cacctggccc ggcaaggcaa cgggccggtg ctggtgtgtg ctccgagcaa
     2401 catcgccgtg gaccagctaa cggagaagat ccaccagacg gggctaaagg tcgtgcgcct
     2461 ctgcgccaag agccgtgagg ccatcgactc cccggtgtct tttctggccc tgcacaacca
     2521 gatcaggaac atggacagca tgcctgagct gcagaagctg cagcagctga aagacgagac
     2581 tggggagctg tcgtctgccg acgagaagcg gtaccgggcc ttgaagcgca ccgcagagag
     2641 agagctgctg atgaacgcag atgtcatctg ctgcacatgt gtgggcgccg gtgacccgag
     2701 gctggccaaa atgcaattta ggagtatttt aatcgacgaa agcacccagg ccaccgagcc
     2761 ggagtgcatg gttcccgtgg tcctcggggc caagcagctg atccttgtag gcgaccactg
     2821 ccagctgggc ccagtggtga tgtgcaagaa ggcggccaag gccgggctgt cacagtcgct
     2881 cttcgagcgc ctggtggtgc tgggcatccg gcccatccgc ctgcaggtcc agtaccggat
     2941 gcaccctgca ctcagcgcct tcccatccaa catcttctac gagggctccc tccagaatgg
     3001 tgtcactgca gcggatcgtg tgaagaaggg atttgacttc cagtggcccc aacccgataa
     3061 accgatgttc ttctacgtga cccagggcca agaggagatt gccagctcgg gcacctccta
     3121 cctgaacagg accgaggctg cgaacgtgga gaagatcacc acgaagttgc tgaaggcagg
     3181 cgccaagccg gaccagattg gcatcatcac gccctacgag ggccagcgct cctacctggt
     3241 gcagtacatg cagttcagcg gctccctgca caccaagctc taccaggagg tggagatcgc
     3301 cagtgtggac gcctttcagg gacgcgagaa ggacttcatc atcctgtcct gtgtgcgggc
     3361 caacgagcac caaggcattg gctttttaaa tgaccccagg cgtctgaacg tggccctgac
     3421 cagagcaagg tatggcgtca tcattgtggg caacccgaag gcactatcaa agcagccgct
     3481 ctggaaccac ctgctgaact actataagga gcagaaggtg ctggtggagg ggccgctcaa
     3541 caacctgcgt gagagcctca tgcagttcag ctaagaattc tgcagtcgac ggtaccgcgg
     3601 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     3661 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3721 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3841 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3901 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3961 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4021 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4081 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4141 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4201 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4261 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4321 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4381 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4441 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4501 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4561 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4621 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4681 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4741 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4801 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4861 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4921 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcacccggtt
     4981 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5041 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5101 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5161 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5221 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5281 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5341 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5401 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5461 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5521 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5581 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5641 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5701 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5761 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5821 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5881 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5941 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6001 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6061 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6121 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6181 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6241 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6301 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6361 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6421 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6481 ctgctgccag tggcgataag tcgtgtctta ctgggttgga ctcaagacga tagttaccgg
     6541 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6601 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6661 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6721 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6781 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6841 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6901 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     6961 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
