LOCUS       sequence-224800-        7989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid driving expression of membrane bound KillerRed fluorophore 
            in zebrafish motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7989)
  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 7989)
  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: pTol2pA2-mnx1:MLS-KillerRed
FEATURES             Location/Qualifiers
     source          1..7989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    2728..2752
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3567..3591)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3593..3727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3790..3810)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3818..3834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3850..3984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(4423..4610,4611..4622)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4656..4672)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4656..4672)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4709..4729)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4709..4727)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4748..4764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4761..4783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4772..4788
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4796..4802,4803..4820,4821..4826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4841..4862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4979..4996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5150..5738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5230..5249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5909..6700,6701..6769))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6532..6551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6770..6874)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7037..7058)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(7407..7418,7419..7581)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     7954..7970
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caactttgta tagaaaagtt gctgaggaaa acgctttcga ctacagaagg actttgtatt
       61 atgcaatgtt ttcaaaaaaa tatcttcata ttcagtgtgg gagataataa gaaaggaaaa
      121 agcatgaagc actgaatcgg tgtcctttaa tgacttttat agttgggaac aattagccgt
      181 tctccacaaa ttaaattgat aaaaagatta cgagatccaa atgcgtttaa aagctgtggt
      241 acgcacggcc catttgtgca attatgatcg aaaatattta ttgcaataaa agagacagtg
      301 caggtcccgt taaattggct tttcataatg aacacattta acatggataa ttaatatata
      361 aactatttcg atttgtcacg atgatatgtg ttgcacttaa ttgtgaaagt aattaccgag
      421 cgaatgaaca tctttgagga gagtccctgg tttcaggccc tcgtatcatc gggtgcaata
      481 ataattttac tctctcctga tgtgatgaaa aagagccata aatctctcag atcgctgcag
      541 gtccacggct tcatatttca ccgctgcttt aatagaaata ttcatgcgat cccttaatga
      601 aattaaaact gagaagcgga caaagcggcc ctaggctatg tgcaacgtgt aaagacacaa
      661 gagcatttga ttgggaaaat gttgcatcct tcaaatatga ggatgttgtt gcactgtaaa
      721 ccacgtaaaa caattttttt cttacagtcc agtattggcc acacttgata attgttacat
      781 ttgttatcaa ttacacgcga ggctcattga atgtcatttc agaattggtc agtgcatgcg
      841 tcattacggc acgcttttat tctcatcctt tcggccggtg acttgttgat ttccctcgag
      901 caaataaacg catgcgacca gtctctcgag agtggttagc tgatgaattg aggaaaacta
      961 atccgcttta ttggtagaca ggttaagggc aacagagtgt caataattct ctccctgacc
     1021 tctttctcca ttagcttaaa gaggcttata agaggaacag accccacaac atgacaccga
     1081 tcctccacat caccatctca catgtagcct acctaggctt ggaaaatcat ttcgatttat
     1141 aaagctcaag atccaacatt tgccaaaaat tcatgaaaac agggcacaaa taggacatca
     1201 ttttctcaca aagtattata taggcctaat tagatagata gatagataga tagatagata
     1261 gatagataga tagatagata gatagataga aaaaccaaga ttttgcttgt gattcattat
     1321 gcataataat ttgcctatgt ttattaataa acatcgtgtt aggattataa atgtgttgtg
     1381 atgaatgtga ttaaatgttt actgacagtt gttagcctat atttacagat gtaaaaaatc
     1441 tacgtctaat tagtatgtgc ttcttaatct ataataagta aaggcttatt aagcttgtct
     1501 ttacaattta gaatttacga atttatcctc cagagaaaaa aaaatgtaat aacaccatat
     1561 ttaaatatcc tgttgagaaa ttatgaaatc caattttcac attctaagac caaggttaaa
     1621 tattgacagt tgacagttgt tttatttttt aagcatgttg tggtgggtga gttttcaaag
     1681 atttacacat ttcaaacaat ttattaaaac aatatttgtt gcataataca ctgatttggt
     1741 cagttttacc actgacttta actaatttat ttaattaatt atatttatat tgttcattgt
     1801 tattaaattt tgcctaccta atgctctcta acaaataagg aataacttga atatttcctc
     1861 tgcgcaaaat gttcatcgta atgtaaaaga tccgtttgac atttaaaatg gttctcttct
     1921 cataaagata gtaattcttt taagaactgt cctctaaaag gttctttggg gaaacaaaaa
     1981 aatatacgtg tacaacatca atgtgaaaat caaatagttt tttttattat cgcaggctgt
     2041 tcagataaaa aaatacaatt gaataaaaaa gtatggacat taaaatggtg tcatagcaaa
     2101 atgacaaatg cgtgtataga ctatggtttt ttgttaatta atttgatgaa ataaaaaaaa
     2161 attatacaaa tgaaaaaaat gtcatgcctc tacatctatg cgaataaaat gcatggctgt
     2221 tttttttttt tgttcaaaca aactatttcg ccaaaataat attttcgagt taacgtgctt
     2281 tatcaatctt cgtttatatt aagattatag gtcggtgttt aaataatgaa atcttgataa
     2341 ctttataaat gtttcatgtt tcaaagttgc atgttaataa ttgcccttta gactctacaa
     2401 atctggaatt attcataaat taagtagatg taaaatgtat gtgtgcatgt tgtgtttccc
     2461 acttaaaaag ctttcatttt atgggagaac atacttatga caactcttga acttctacaa
     2521 gcacaccgtg aacactggaa tcactgtcca atcggaaatg cacaagaatg acgtgaatga
     2581 ccaatccact ccctctcggg cagcagttga caagtggact tctggcttgc acaccttggc
     2641 caagatcata actttacgaa gaggagcggt gacactcctg acagcctttt aaagttacga
     2701 acgacgcgtt aatctgtttg tgaggtggca agtttgtaca aaaaagcagg cttacgggat
     2761 cgatccaccg gtcgccacca tgctgtgctg tatgagaaga accaaacagg ttgaaaagaa
     2821 tgatgaggac caaaagatct ccgagggcgg ccccgccctg ttccagagcg acatgacctt
     2881 caaaatcttc atcgacggcg aggtgaacgg ccagaagttc accatcgtgg ccgacggcag
     2941 cagcaagttc ccccacggcg acttcaacgt gcacgccgtg tgcgagaccg gcaagctgcc
     3001 catgagctgg aagcccatct gccacctgat ccagtacggc gagcccttct tcgcccgcta
     3061 ccccgacggc atcagccatt tcgcccagga gtgcttcccc gagggcctga gcatcgaccg
     3121 caccgtgcgc ttcgagaacg acggcaccat gaccagccac cacacctacg agctggacga
     3181 cacctgcgtg gtgagccgca tcaccgtgaa ctgcgacggc ttccagcccg acggccccat
     3241 catgcgcgac cagctggtgg acatcctgcc caacgagacc cacatgttcc cccacggccc
     3301 caacgccgtg cgccagctgg ccttcatcgg cttcaccacc gccgacggcg gcctgatgat
     3361 gggccacttc gacagcaaga tgaccttcaa cggcagccgc gccatcgaga tccccggccc
     3421 acacttcgtg accatcatca ccaagcagat gagggacacc agcgacaagc gcgaccacgt
     3481 gtgccagcgc gaggtggcct acgcccacag cgtgccccgc atcaccagcg ccatcggtag
     3541 cgacgaggat taaagcggcc aattctaccc agctttcttg tacaaagtgg gggatccaga
     3601 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3661 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3721 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3781 ggttttttcc aactttatta tacatagttg ataattcact ggccgtcgtt ttacggtacc
     3841 atcgatgatg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3901 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3961 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4021 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     4081 gattatgatc ctctagatca gatctgcgaa gatacggcca cgggtgctct tgatcctgtg
     4141 gctgattttg gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct
     4201 tccactgatt gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt
     4261 aatagatgtg acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata
     4321 taatgtgaaa acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag
     4381 cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat
     4441 aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa
     4501 aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat
     4561 ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc
     4621 tggttcttga ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc
     4681 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     4741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5341 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5581 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5701 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5761 cggggtctga cgctcagtgg aacgaaaact cacgttaagg cattttggtc atgagattat
     5821 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     6901 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     6961 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     7021 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7081 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca cgctggctaa gaactcatca
     7141 gcctccccgg tccatctacc cacgtaccaa tgcaccaatt ggccacaatg acggctacta
     7201 catggtgcca ttccttcctc tttataggaa tggagactac ctcctgtcca acaatgctct
     7261 tggatacgag tacgcctacc tgttggaccc aggtcattgc acaacaccag aaatgccctc
     7321 tgatctgcaa aagacgtgaa tatctgttca gacacccata tccactctgt tccacacagg
     7381 tcagaggttt gtccaggagt tcttgacaga ggtgtaaaaa gtactcaaaa attttactca
     7441 agtgaaagta caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta
     7501 gaatcttact tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaagg
     7561 aagtaaaagt aaaagcaaga aagaaaacta gagattcttg tttaagcttt taatctcaaa
     7621 aaacattaaa tgaaatgcat acaaggtttt atcctgcttt agaactgttt gtatttaatt
     7681 atcaaactat aagacagaca atctaatgcc agtacacgct actcaaagtt gtaaaacctc
     7741 agatttaact tcagtagaag ctgattctca aaattgttag tgtcaagcct agctcttttg
     7801 gggctgaaaa gcaatcctgc agtgctgaaa agcctctcac aggcagccga tgcgggaaga
     7861 ggtgtattag tcttgataga gaggctgcaa atagcaggaa acgtgagcag agactccctg
     7921 gtgtctgaaa cacaggccag atgggccctc gagcaggaaa cagctatgac catgattacg
     7981 ccaagctat
//
