LOCUS       sequence-217834-        3354 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 3354)
  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 3354)
  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..3354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(7..106)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(123..142)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(124..142)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(147..163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(147..163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(265..284)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             276..1085
                     /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     complement(343..362)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2150..2177)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2269..2355)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2404..2426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2418..2435
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2419..2435
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2451..2550
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aattctaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
       61 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
      121 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
      181 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
      241 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
      301 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
      361 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
      421 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
      481 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
      541 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
      601 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
      661 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
      721 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
      781 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
      841 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
      901 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
      961 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     1021 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     1081 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     1141 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     2161 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     2221 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     2281 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     2341 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     2401 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga
     2461 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta
     2521 taatgccaac tttgtacaaa aaagcaggct tacgggatcg atccaccggt cgccaccatg
     2581 ctgtgctgta tgagaagaac caaacaggtt gaaaagaatg atgaggacca aaagatctcc
     2641 gagggcggcc ccgccctgtt ccagagcgac atgaccttca aaatcttcat cgacggcgag
     2701 gtgaacggcc agaagttcac catcgtggcc gacggcagca gcaagttccc ccacggcgac
     2761 ttcaacgtgc acgccgtgtg cgagaccggc aagctgccca tgagctggaa gcccatctgc
     2821 cacctgatcc agtacggcga gcccttcttc gcccgctacc ccgacggcat cagccatttc
     2881 gcccaggagt gcttccccga gggcctgagc atcgaccgca ccgtgcgctt cgagaacgac
     2941 ggcaccatga ccagccacca cacctacgag ctggacgaca cctgcgtggt gagccgcatc
     3001 accgtgaact gcgacggctt ccagcccgac ggccccatca tgcgcgacca gctggtggac
     3061 atcctgccca acgagaccca catgttcccc cacggcccca acgccgtgcg ccagctggcc
     3121 ttcatcggct tcaccaccgc cgacggcggc ctgatgatgg gccacttcga cagcaagatg
     3181 accttcaacg gcagccgcgc catcgagatc cccggcccac acttcgtgac catcatcacc
     3241 aagcagatga gggacaccag cgacaagcgc gaccacgtgt gccagcgcga ggtggcctac
     3301 gcccacagcg tgccccgcat caccagcgcc atcggtagcg acgaggatta aagc
//
