LOCUS       sequence-223591-        3358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry plasmid containing membrane bound KillerRed 
            fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3358)
  AUTHORS   Formella I, Svahn AJ, Radford RAW, Don EK, Cole NJ, Hogan A, Lee A, 
            Chung RS, Morsch M
  TITLE     Real-time visualization of oxidative stress-mediated 
            neurodegeneration of individual spinal motor neurons in vivo.
  JOURNAL   Redox Biol. 2018 Aug 23;19:226-234. doi: 
            10.1016/j.redox.2018.08.011.
  PUBMED    30193184
REFERENCE   2  (bases 1 to 3358)
  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: "Redox 
            Biol."; date: "2018-08-23"; volume: "19:226-234. doi"; pages: " 
            10.1016/j.redox.2018.08.011"
COMMENT     Sequence Label: pME-MLS-KillerRed
FEATURES             Location/Qualifiers
     source          1..3358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(915..1014)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1030..1047)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1030..1046)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1039..1061)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1110..1196
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1288..1315
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1474..1491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1645..2233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1725..1744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2380..3189)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3103..3122
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3181..3200
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3302..3318
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3302..3318
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3323..3342
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3323..3341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa
       61 tgctttctta taatgccaac tttgtacaag aaagctgggt agaattggcc gctttaatcc
      121 tcgtcgctac cgatggcgct ggtgatgcgg ggcacgctgt gggcgtaggc cacctcgcgc
      181 tggcacacgt ggtcgcgctt gtcgctggtg tccctcatct gcttggtgat gatggtcacg
      241 aagtgtgggc cggggatctc gatggcgcgg ctgccgttga aggtcatctt gctgtcgaag
      301 tggcccatca tcaggccgcc gtcggcggtg gtgaagccga tgaaggccag ctggcgcacg
      361 gcgttggggc cgtgggggaa catgtgggtc tcgttgggca ggatgtccac cagctggtcg
      421 cgcatgatgg ggccgtcggg ctggaagccg tcgcagttca cggtgatgcg gctcaccacg
      481 caggtgtcgt ccagctcgta ggtgtggtgg ctggtcatgg tgccgtcgtt ctcgaagcgc
      541 acggtgcggt cgatgctcag gccctcgggg aagcactcct gggcgaaatg gctgatgccg
      601 tcggggtagc gggcgaagaa gggctcgccg tactggatca ggtggcagat gggcttccag
      661 ctcatgggca gcttgccggt ctcgcacacg gcgtgcacgt tgaagtcgcc gtgggggaac
      721 ttgctgctgc cgtcggccac gatggtgaac ttctggccgt tcacctcgcc gtcgatgaag
      781 attttgaagg tcatgtcgct ctggaacagg gcggggccgc cctcggagat cttttggtcc
      841 tcatcattct tttcaacctg tttggttctt ctcatacagc acagcatggt ggcgaccggt
      901 ggatcgatcc cgtaagcctg cttttttgta caaagttggc attataaaaa agcattgctc
      961 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
     1021 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt
     1081 taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
     1141 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1201 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1261 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
     1321 ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cactgcccgc
     1381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2281 cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt
     2341 aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat
     2401 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     2461 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     2521 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     2581 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     2641 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     2701 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     2761 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     2821 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     2881 tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     2941 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     3001 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3061 cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     3121 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat
     3181 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     3241 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccagagctg
     3301 ccaggaaaca gctatgacca tgtaatacga ctcactatag gggatatcag ctggatgg
//
