LOCUS       sequence-330245-        4656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N. furzeri MARK4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4656)
  AUTHORS   Chen YR, Harel I, Singh PP, Ziv I, Moses E, Goshtchevsky U, Machado 
            BE, Brunet A, Jarosz DF
  TITLE     Tissue-specific landscape of protein aggregation and quality control
            in an aging vertebrate.
  JOURNAL   Dev Cell. 2024 Jul 22;59(14):1892-1911.e13. doi: 
            10.1016/j.devcel.2024.04.014. Epub 2024 May 28.
  PUBMED    38810654
REFERENCE   2  (bases 1 to 4656)
  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: 
            "10.1016/j.devcel.2024.04.014"; journalName: "Dev Cell"; date: 
            "2024-07-22- 22"; volume: "59"; issue: "14"; pages: "1892-1911.e13"
COMMENT     Sequence Label: pDONR221-MARK4(v1)
FEATURES             Location/Qualifiers
     source          1..4656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2781..2880)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2897..2916)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2898..2916)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2921..2937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2921..2937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3039..3058)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3050..3859
                     /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(3117..3136)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4006..4594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4495..4514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc gaaggagata acaaaatgtc ctctcgatct gcgttaccgt ctgggagcga
      721 caggagcaca ggggaacatc atgtgtcgct gggggccagc cgttcagaca aggccacgag
      781 ccagtccagc cgctctctgg gcgcccgctg caggaactcc atcgcctcct gctctgatga
      841 gcagccacac attggcaact accgtctgct caagaccatt ggcaagggaa actttgccaa
      901 agtcaaactg gctcgccaca ttctgacagg cagagaggtt gcaataaaaa tcatcgataa
      961 aactcagttg aacccaacca gcctacaaaa gcttttccgt gaggtgcgaa taatgaaggg
     1021 cctaaatcac cctaacatag tgaagctgtt tgaggtgatt gagacggata agacccttta
     1081 cctggtcatg gagtatgcca gtggagggga agtgtttgac tacctcgtgt ctcatgggag
     1141 aatgaaggag gttgaagcca gagccaaatt tcgtcagatc gtctctgctg tccactactg
     1201 ccacaccaag aacatagtcc accgagatct gaaggcggag aaccttctgc tggatgccga
     1261 cgctaacatc aagatagctg attttggctt cagcaatgag ttcactctgg gaaacaagct
     1321 ggacacgttc tgtggctcgc tgccctacgc tgcccccgag ctcttccagg ggaagaagta
     1381 tgatgggccc gaggtggatg tctggagtct gggagtcatc ctctacactc tggtcagcgg
     1441 ctcgctgcct tttgatggac agaacctcaa ggagctgagg gagcgtgtgc tgagaggaaa
     1501 gtaccgagtg cccttctaca tgtcaactga ctgtgaggga atccttcgcc gattcctggt
     1561 gctcaatcca gccaagcgct gcacgctgga gcaagtcatg aaggacaaat ggataaacac
     1621 agggtacgag ggtgacgacc tgaagcccca catagagcct gtggaggact acagtgatcc
     1681 ggctcgcata gaggtgatgg tcgggatggg cttcaccccc gaggaaatca aagactctct
     1741 gctcaatcag aaatacaacg aggtcaccgc cacatatttg ctcctgggcc gaaaaggcga
     1801 tgagagcagt gatgcccgct ctgctagtag cctgtccttg gcgagggtac gacccagccc
     1861 gatcactaac gggaccaaca aacactcctc agccactggc tcctcctcct cctccacctc
     1921 ctcttcgcat agcaagaccc agcgcagtgc ctccacctac cacaggcagc gacggcacag
     1981 cgacttctgt gggccttcca tgcccgtcat gcaccccaag cggagcccga ccagcacggg
     2041 ggaccgggag ctgcgggaat gccggatgcc ttctcgtaag gccagctgca gcgttatggg
     2101 gagccacagc ctgcccccct ccagcccaat ggtcagcagt gccaataacc ccaacaagtc
     2161 cgagatccca gaccgccgca aagacatgac tgccaccacg aataacatcc ctgggagcac
     2221 catgacacgc cggaacacct acgtgtgcac ggaccgctcg ggcgctgaca gacactcact
     2281 cctgcagaat ggcaaagaaa acagctctct gggtcaccgc ctgcctgcag cttctccctc
     2341 cacgctcagc atctctggag ctggagcagc ttcctcctcg tcctcttcgg accggtgccg
     2401 tttgacccga gggtccacca tccgcagcac cttccatggg ggacagctga gagaccgtgg
     2461 tcctcccgtc tactctgccc cacccacatc acccaccctg tcccaccatg aagccagccc
     2521 gttgccccat gctcgcacca gggcaacctc caacctcttc accaagctga cctcaaaact
     2581 cactcgcagg gtcaacctag acccgtccaa gcgccaaggc tcaaacaaat cagtctcggg
     2641 ttgtactctt ccacaaggat caaaaacagt taggtcacaa atgaatctga gggaatcagg
     2701 agacttgcgg tcacaagttg ccatctacct gggtatcaaa aagcgtccga gccccgggcc
     2761 atcggacgtg gctggaatcg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2821 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2881 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2941 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3001 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3061 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3121 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3181 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3241 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3301 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3361 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3421 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3481 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3541 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3601 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3661 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3721 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3781 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3841 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3901 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3961 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4021 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4081 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4141 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4201 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4261 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4321 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4381 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4441 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4501 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4561 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4621 gttcctggcc ttttgctggc cttttgctca catgtt
//
