LOCUS       sequence-245492-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses peroxisomal KillerRed (po-KR) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4683)
  AUTHORS   Ivashchenko O, Van Veldhoven PP, Brees C, Ho YS, Terlecky SR, 
            Fransen M
  TITLE     Intraperoxisomal redox balance in mammalian cells: oxidative stress 
            and interorganellar cross-talk.
  JOURNAL   Mol Biol Cell. 2011 May;22(9):1440-51. doi: 10.1091/mbc.E10-11-0919.
            Epub 2011 Mar 3.
  PUBMED    21372177
REFERENCE   2  (bases 1 to 4683)
  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: 
            "10.1091/mbc.E10-11-0919"; journalName: "Mol Biol Cell"; date: 
            "2011-05"; volume: "22"; issue: "9"; pages: "1440-51"
COMMENT     Sequence Label: pMF1754
FEATURES             Location/Qualifiers
     source          1..4683
                     /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"
     CDS             join(558..560,561..566,567..1274)
                     /codon_start=1
                     /product="red fluorescent protein KillerRed, a 
                     genetically-encoded photosensitizer (Bulina et al., 2006)"
                     /label=KillerRed
                     /note="mammalian codon-optimized"
                     /translation="MGSEGGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDDTCVVSRITVNCDGFQPDGPIMRDQLVDILPNETHMFPHGPNAVRQLA
                     FIGFTTADGGLMMGHFDSKMTFNGSRAIEIPGPHFVTIITKQMRDTSDKRDHVCQREVA
                     YAHSVPRITSAIGSDED"
     polyA_signal    1411..1532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1448..1467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1502..1521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1539..1994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1676..1697)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1888..1907
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2021..2125
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2127..2484
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2127..2318
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2150..2170)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2335..2470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2519..3313
                     /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             2519..3313
                     /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(2573..2592)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3183..3202
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3501..3520)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3545..3592
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3921..4509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4410..4429
                     /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 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctaccatg ggttcagagg gcggccccgc cctgttccag agcgacatga
      601 ccttcaaaat cttcatcgac ggcgaggtga acggccagaa gttcaccatc gtggccgacg
      661 gcagcagcaa gttcccccac ggcgacttca acgtgcacgc cgtgtgcgag accggcaagc
      721 tgcccatgag ctggaagccc atctgccacc tgatccagta cggcgagccc ttcttcgccc
      781 gctaccccga cggcatcagc catttcgccc aggagtgctt ccccgagggc ctgagcatcg
      841 accgcaccgt gcgcttcgag aacgacggca ccatgaccag ccaccacacc tacgagctgg
      901 acgacacctg cgtggtgagc cgcatcaccg tgaactgcga cggcttccag cccgacggcc
      961 ccatcatgcg cgaccagctg gtggacatcc tgcccaacga gacccacatg ttcccccacg
     1021 gccccaacgc cgtgcgccag ctggccttca tcggcttcac caccgccgac ggcggcctga
     1081 tgatgggcca cttcgacagc aagatgacct tcaacggcag ccgcgccatc gagatccccg
     1141 gcccacactt cgtgaccatc atcaccaagc agatgaggga caccagcgac aagcgcgacc
     1201 acgtgtgcca gcgcgaggtg gcctacgccc acagcgtgcc ccgcatcacc agcgccatcg
     1261 gtagcgacga ggataaaagc aagctgtgag cggccgcgac tctagatcat aatcagccat
     1321 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1381 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1441 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1501 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     1561 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1621 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1681 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1741 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1801 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1861 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1921 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1981 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2101 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     2161 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2221 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2281 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2341 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2401 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2461 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     2521 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2581 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2641 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2701 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2761 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2821 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2881 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2941 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3001 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     3061 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3121 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3181 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3241 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3301 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3361 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3421 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3481 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3541 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     3601 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3661 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3721 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     3781 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3841 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3901 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3961 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4021 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4081 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4141 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4201 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4261 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4321 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4381 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4441 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4501 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4561 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     4621 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     4681 ccg
//
