LOCUS       sequence-328876-        3276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3276)
  AUTHORS   Harel I, Chen YR, Ziv I, Singh PP, Heinzer D, Navarro Negredo P, 
            Goshtchevsky U, Wang W, Astre G, Moses E, McKay A, Machado BE, 
            Hebestreit K, Yin S, Sanchez Alvarado A, Jarosz DF, Brunet A
  TITLE     Identification of protein aggregates in the aging vertebrate brain 
            with prion-like and phase-separation properties.
  JOURNAL   Cell Rep. 2024 Jun 25;43(6):112787. doi: 
            10.1016/j.celrep.2023.112787. Epub 2024 May 28.
  PUBMED    38810650
REFERENCE   2  (bases 1 to 3276)
  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: "Cell Rep.";
            date: "2024-06-25"; volume: "43(6):112787. doi"; pages: " 
            10.1016/j.celrep.2023.112787. Epub 2024 May 28"
COMMENT     Sequence Label: pDONR221-mRuby3
FEATURES             Location/Qualifiers
     source          1..3276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(60..146)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     195..217
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     209..226
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     210..226
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    242..341
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             359..1069
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     protein_bind    complement(1074..1173)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1190..1209)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1191..1209)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1214..1230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1214..1230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1332..1351)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1343..2152
                     /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(1410..1429)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2299..2887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2788..2807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3041..3058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3217..3244)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
       61 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      121 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      181 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      241 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
      301 atgctttttt ataatgccaa ctttgtacaa aaaagcaggc ttcgaaggag ataacaaaat
      361 ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg
      421 ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcagac cgtacgaggg
      481 agtgcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat
      541 tcttgccacg tcgttcatgt atggcagccg tacctttatc aagtacccgg ccgacatccc
      601 tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga
      661 agatggtgga gtcgtcaccg tcacgcagga caccagcctt gaggatggcg agctcgtcta
      721 caacgtcaag gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac
      781 caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc tgagaggata
      841 cactgacatc gcactgaaag ttgatggtgg tggccatctg cactgcaact tcgtgacaac
      901 ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtgtccatg ccgttgatca
      961 ccgcctggaa aggatcgagg agagtgacaa tgaaacctac gtagtgcaaa gagaagtggc
     1021 agttgccaaa tacagcaacc ttggtggtgg catggacgag ctgtacaagt aagacccagc
     1081 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1141 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1201 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1261 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1321 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1381 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1441 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1501 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1561 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1621 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1681 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1741 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1801 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     1861 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     1921 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     1981 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2041 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2101 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2161 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2221 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2281 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2341 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2401 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2461 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2521 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2581 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2641 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2701 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2761 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2821 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2881 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2941 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3001 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3061 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3121 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     3181 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3241 ttctgcttag tttgatgcct ggcagtttat ggcggg
//
