LOCUS       sequence-414755-        4513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PuRRRE genetically-encoded tag introduced after TOMM20; to be used 
            with PunyEEER probe peptide..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4513)
  AUTHORS   Suyama A, Devlin KL, Macias-Contreras M, Doh JK, Shinde U, Beatty KE
  TITLE     Orthogonal Versatile Interacting Peptide Tags for Imaging Cellular 
            Proteins.
  JOURNAL   Biochemistry. 2023 Jun 6;62(11):1735-1743. doi: 
            10.1021/acs.biochem.2c00712. Epub 2023 May 11.
  PUBMED    37167569
REFERENCE   2  (bases 1 to 4513)
  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"; doi: 
            "10.1021/acs.biochem.2c00712"; journalName: "Biochemistry"; date: 
            "2023-06-6- 6"; volume: "62"; issue: "11"; pages: "1735-1743"
COMMENT     Sequence Label: TOMM20-PuRRRE
FEATURES             Location/Qualifiers
     source          1..4513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..258
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     215..235
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(260..364)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      391..846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(478..497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     688..709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    853..974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(864..883)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     918..937
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      1669..1678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1707..1910)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1736..1756)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1911..2214
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2389..2977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2469..2488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3306..3353
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3378..3397
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3585..4379)
                     /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             complement(3585..4379)
                     /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(3696..3715)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4306..4325
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4414..4513,1..258))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4428..4513,1..50)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
       61 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      121 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      181 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacagc
      241 tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      361 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
      421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      541 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
      721 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
      781 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
      841 caatttacgc cttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      901 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      961 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1021 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     1081 tagagtcgcg gccgctttac agcggaccgt attcgttttc cagttcggag ttttcctgac
     1141 gcagttcaga aacgcgggta cgcagcgcgg taacacgctg acggaggaag gcgttacgga
     1201 tttccagggt ggcgaccggt ggatccccgc taccgccttc cacatcatct tcagccaagc
     1261 tctgagcact tacaattctc tgactaattg ttgggagctt agtcagaagc atctggaaca
     1321 ctggtggtgg aagagtttgc tgtaagacct gcagtaactg ctgtggctgt ccacacacag
     1381 caattgcatt tgtcagatgg tctacgccct tctcatattc accttgagct agtaactctt
     1441 caccaagctg tatttcttca aggaagaact tctgaacagc ttcagcatct ttaaggtcag
     1501 gtaacttgga aagcccagct ctctccttgg caagcttctg tttctttctt cgttctcgaa
     1561 gcctgttctt gaagttgggg tcacttcgtc ttttgcggtc gaagtagatg cagtacccaa
     1621 tgaaaagggc cccgcataca ccggcggcga tggcgctgtt ccgacccacc atggtggcgc
     1681 tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc
     1741 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     1801 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     1861 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     1921 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     1981 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     2041 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     2101 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     2161 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     2221 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actaatgcat
     2281 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2341 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2401 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2461 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2521 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2581 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2641 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2701 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2761 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2821 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2881 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2941 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3001 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3061 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3121 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     3181 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     3241 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     3301 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     3361 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     3421 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     3481 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     3541 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     3601 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     3661 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     3721 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     3781 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     3841 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     3901 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     3961 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     4021 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     4081 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     4141 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     4201 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4261 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4321 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     4381 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     4441 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     4501 aaaaaaaatt agt
//
