LOCUS       sequence-430443-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUpf1-del296-926.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5726)
  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-del296-926_P
FEATURES             Location/Qualifiers
     source          1..5726
                     /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    2454..2575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2491..2510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2545..2564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2582..3037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2719..2740)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2931..2950
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3064..3168
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3170..3527
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3170..3361
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3193..3213)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3378..3513
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3562..4356
                     /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             3562..4356
                     /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(3616..3635)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4226..4245
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4544..4563)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4588..4635
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4964..5552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5453..5472
                     /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 gccacggaag ctggtcaaca ctatcaaccc gggagcccgc ttcatgacca cagccatgta
     1741 tgatgcccgg gaggccatca tcccaggctc cgtctatgat cggagcagcc agggccggcc
     1801 ttccagcatg tacttccaga cccatgacca gattggcatg atcagtgccg gccctagcca
     1861 cgtggctgcc atgaacattc ccatcccctt caacctggtc atgccaccca tgccaccgcc
     1921 tggctatttt ggacaagcca acgggcctgc tgcagggcga ggcaccccga aaggcaagac
     1981 tggtcgtggg ggacgccaga agaaccgctt tgggcttcct ggacccagcc agactaacct
     2041 ccccaacagc caagccagcc aggatgtggc gtcacagccc ttctctcagg gcgccctgac
     2101 gcagggctac atctccatga gccagccttc ccagatgagc cagcccggcc tctcccagcc
     2161 ggagctgtcc caggacagtt accttggtga cgagtttaaa tcacaaatcg acgtggcgct
     2221 ctcacaggac tccacgtacc agggagagcg ggcttaccag catggcgggg tgatggggct
     2281 gtcccagtat taagaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     2341 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2401 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2461 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2521 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2581 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2641 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2701 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2761 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2821 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2881 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2941 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3001 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3061 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3121 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3181 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3421 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3541 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3601 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3661 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3721 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3781 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3841 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3901 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3961 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4021 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4081 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4141 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4201 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4261 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4321 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4381 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4441 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4501 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4561 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4621 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4681 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4741 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4801 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4861 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4921 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4981 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5041 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5101 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5161 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5221 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5281 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5341 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5401 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5461 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5521 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5581 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5641 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
     5701 gttcatagcc catatatgga gttccg
//
