LOCUS       sequence-348949-        3518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Site-specific mutagenesis of mOrange for the purposes of creating a 
            site-specific 8-oxo-G:C lesion for the purposes of measuring OGG1 
            activity via Host Cell Reactivation (HCR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  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 3518)
  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_mOrange_A215C
FEATURES             Location/Qualifiers
     source          1..3518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          676..808
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      825..834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             831..1541
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTSGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1244..1263
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1554..1573)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    1583..1664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1608..1627
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(1862..2450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1942..1961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2531..3325)
                     /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(2531..3325)
                     /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(2642..2661)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3252..3271
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cctgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aggtcgttta gtgaaccgtc
      601 agatcactag tagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      661 ctcgactgat cacaggtaag tatcaaggtt acaagacagg tttaaggagg ccaatagaaa
      721 ctgggcttgt cgagacagag aagattcttg cgtttctgat aggcacctat tggtcttact
      781 gacatccact ttgcctttct ctccacaggg gtaccgctag cgctgccacc atggtgagca
      841 agggcgagga gaataacatg gccatcatca aggagttcat gcgcttcaag gtgcgcatgg
      901 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
      961 agggctttca gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     1021 acatcctgtc ccctcagttc acctccggct ccaaggccta cgtgaagcac cccgccgaca
     1081 tccccgacta cttcaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1141 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     1201 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     1261 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     1321 gcgagatcaa gatgaggctg aagctgaagg acggcggcca ctacacctcc gaggtcaaga
     1381 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc ggcatcaagt
     1441 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     1501 gccgccactc caccggcggc atggacgagc tgtacaagta gaagcttgag ctcgatgagt
     1561 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1621 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1681 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1741 tctacaaatg tggtacttaa gagggggaga ccaaagggcg agacgttaag gcctcacgtg
     1801 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1861 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1921 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1981 gctctcctgt tccgaccctg ccgcttacgg gatacctgtc cgcctttctc ccttcgggaa
     2041 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2101 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2161 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2221 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2281 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2341 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2401 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2461 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2521 tcatgccgtc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     2581 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     2641 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     2701 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     2761 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     2821 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     2881 ccatccgagt acgtgctctc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     2941 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3001 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3061 ttcccgcttc agtgacaacg tcgagtacag ctgcgcaagg aacgcccgtc gtggccagcc
     3121 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     3181 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3241 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3301 cgtgcaatcc atcttgttca atcataatat tattgaagca tttatcaggg ttcgtctcgt
     3361 cccggtctcc tcccatgcat gtcaatattg gccattagcc atattattca ttggttatat
     3421 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     3481 catttatatt ggctcatgtc caatatgacc gccatgtt
//
