LOCUS       sequence-109465-        5111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressses V5- and SBP-tagged RRP4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5111)
  TITLE     Izaurralde lab Exosc plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5111)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pT7-V5-SBP-C1-HsRRP4
FEATURES             Location/Qualifiers
     source          1..5111
                     /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    1899..2020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1936..1955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1990..2009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2027..2482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2164..2185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2376..2395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2509..2613
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2615..2972
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2615..2806
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2638..2658)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2823..2958
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3007..3801
                     /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             3007..3801
                     /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(3061..3080)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3671..3690
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3989..4008)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4033..4080
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4409..4997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4898..4917
                     /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 cgagggatgg cgatggagat gaggcttcca gtggctcgca agcctcttag
      901 cgagagactg ggccgcgaca ctaagaaaca tctagtggtg ccgggggata caatcactac
      961 ggacacagga ttcatgcggg gccatggaac gtatatggga gaagagaagc tcattgcatc
     1021 tgttgctggc tctgtggaga gagtaaacaa gttgatctgt gtgaaagctt tgaaaaccag
     1081 atacattggt gaagtaggag acatcgtagt gggacgaatc acagaggttc aacagaagag
     1141 gtggaaggtg gagaccaact ccaggctgga ttcggtcttg ctgctctcgt ccatgaacct
     1201 tcctggagga gagctgagga gaagatctgc agaagatgag cttgcaatga gaggtttctt
     1261 acaggaaggg gaccttatca gtgctgaggt ccaggcagtg ttctctgacg gagctgtctc
     1321 tttgcacacg aggagcctga aatatggaaa actaggtcag ggggttttgg tccaggtttc
     1381 cccctccctg gtgaaacggc agaagaccca ctttcatgat ttgccatgtg gtgcctcagt
     1441 gattctcggt aacaacggct tcatctggat ttacccaaca cctgagcaca aagaagagga
     1501 agcagggggc ttcattgcaa acctggagcc tgtctctctt gctgatcgag aggtgatatc
     1561 ccggcttcgg aactgcatca tctcgctggt aactcagagg atgatgctgt atgataccag
     1621 catcctgtac tgctatgaag catcccttcc acatcagatc aaagacatct taaagccaga
     1681 aataatggag gagattgtga tggaaacacg ccagaggctt ttggaacagg agggataaga
     1741 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     1801 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1861 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1981 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2041 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2101 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2161 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2221 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2281 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2341 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2401 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2461 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2521 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2581 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2641 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2761 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2821 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2881 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2941 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3001 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3061 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3121 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3181 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3241 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3301 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3361 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3421 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3481 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3541 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3601 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3661 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3721 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3781 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3841 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3901 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3961 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4021 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4081 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4141 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4201 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4261 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4321 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4381 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4441 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4501 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4561 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4621 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4681 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4741 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4801 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4861 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4921 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4981 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5041 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5101 ccgccatgca t
//
