LOCUS       sequence-348952-        3503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Site-specific mutagenesis of tagBFP for the purposes of creating a 
            site-specific U:A mispair for the purposes of measuring UDG activity
            via Host Cell Reactivation (HCR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3503)
  AUTHORS   Nagel ZD, Margulies CM, Chaim IA, McRee SK, Mazzucato P, Ahmad A, 
            Abo RP, Butty VL, Forget AL, Samson LD
  TITLE     Multiplexed DNA repair assays for multiple lesions and multiple 
            doses via transcription inhibition and transcriptional mutagenesis.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 May 6;111(18):E1823-32. doi: 
            10.1073/pnas.1401182111. Epub 2014 Apr 22.
  PUBMED    24757057
REFERENCE   2  (bases 1 to 3503)
  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: 
            "10.1073/pnas.1401182111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-05-6- 6"; volume: "111"; issue: "18"; pages: "E1823-32"
COMMENT     Sequence Label: pMax_BFP_A191G
FEATURES             Location/Qualifiers
     source          1..3503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          296..428
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             451..1152
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLCGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1159..1178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    1188..1269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1213..1232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(1467..2055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1547..1566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2136..2930)
                     /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(2136..2930)
                     /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(2247..2266)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2857..2876
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     enhancer        3124..3503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag aggtcgttta gtgaaccgtc agatcactag tagctttatt
      241 gcggtagttt atcacagtta aattgctaac gcagtcagtg ctcgactgat cacaggtaag
      301 tatcaaggtt acaagacagg tttaaggagg ccaatagaaa ctgggcttgt cgagacagag
      361 aagattcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
      421 ctccacaggg gtaccgctag cgctgccacc atgagcgagc tgattaagga gaacatgcac
      481 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
      541 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
      601 ctccccttcg ccttcgacat cctggctact agcttcctct gcggcagcaa gaccttcatc
      661 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
      721 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
      781 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
      841 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
      901 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
      961 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
     1021 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
     1081 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
     1141 aagcttaatt gagagctcga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     1201 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1261 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1321 gaggtttttt aaagcaagta aaacctctac aaatgtggta cttaagaggg ggagaccaaa
     1381 gggcgagacg ttaaggcctc acgtgacatg tgagcaaaag gccagcaaaa ggccaggaac
     1441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct tacgggatac
     1621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2101 gaaaactcac gttaagggat tttggtcatg ccgtctcaga agaactcgtc aagaaggcga
     2161 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     2221 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     2281 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     2341 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     2401 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     2461 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctctctcgat gcgatgtttc
     2521 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     2581 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     2641 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag tacagctgcg
     2701 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     2761 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     2821 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     2881 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat aatattattg
     2941 aagcatttat cagggttcgt ctcgtcccgg tctcctccca tgcatgtcaa tattggccat
     3001 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     3061 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
     3121 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3361 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
     3421 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
     3481 tattagtcat cgctattacc atg
//
