LOCUS       sequence-328993-        3204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N. furzeri CALM3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3204)
  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 3204)
  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-CALM3(3of4)
FEATURES             Location/Qualifiers
     source          1..3204
                     /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"
     CDS             881..1327
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     protein_bind    complement(1329..1428)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1445..1464)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1446..1464)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1469..1485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1469..1485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1587..1606)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1598..2407
                     /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(1665..1684)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2554..3142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3043..3062
                     /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 acaaaatgct ctcgtgctgg gggcggggct gctgctgcaa
      721 gccggaatac ggaaatgagt gcggcggagg gatacacaca caagccgacc aatatacaca
      781 gtcggcttgt gtgtacgccg ccgccagtgg aacgccattt gagggacgag ctcacttcca
      841 gactgcctcc acggaccgac aagcatacaa cccgtccgaa atggctgacc aattaacaga
      901 ggaacagatc gcagagttta aagaggcttt ctccttattc gacaaagacg gcgacggaac
      961 catcaccacc aaggagcttg ggacggtcat gaggtcgctg ggtcagaacc ccacagaggc
     1021 agaacttcag gatatgatta atgaggtgga tgctgacggt aacggaacca tcgacttccc
     1081 tgagtttttg accatgatgg caaggaaaat gaaggacaca gacagcgagg aggagatccg
     1141 cgaggctttc cgggtattcg acaaggacgg aaacggatac attagtgctg cggagctgcg
     1201 tcacgtcatg acgaacctag gagagaagct aacggacgag gaggtggacg agatgatcag
     1261 agaagcagac atcgacgggg atggacaggt caactacgaa gagtttgtac agatgatgac
     1321 tgcgaaggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1381 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1441 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1501 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1561 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1621 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1681 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1741 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1801 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1861 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1921 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1981 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2041 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2101 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2161 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2221 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2281 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2341 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2401 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2461 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3181 ttgctggcct tttgctcaca tgtt
//
