LOCUS       sequence-317994-        3524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-fragment Gateway compatible flourophore for expression in C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3524)
  AUTHORS   Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM, 
            Frokjaer-Jensen C
  TITLE     Engineering rules that minimize germline silencing of transgenes in 
            simple extrachromosomal arrays in C. elegans.
  JOURNAL   Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.
  PUBMED    33298957
REFERENCE   2  (bases 1 to 3524)
  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: "Nat 
            Commun."; date: "2020-12-9"; volume: "11(1):6300. doi"; pages: " 
            10.1038/s41467-020-19898-0"
COMMENT     Sequence Label: pCFJ2245 - [1-2] ENTR - Fluor - ce-tagBFP2 (1
            ...
FEATURES             Location/Qualifiers
     source          1..3524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1113..1212)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1229..1248)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1230..1248)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1253..1269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1253..1269)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1371..1390)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1382..2191
                     /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(1449..1468)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2338..2926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2827..2846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3080..3097
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3256..3283)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3375..3461)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3510..3524,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3524,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctcggccggc agtactagcg gtggcagtgg aggtaccggc ggaagcagtt
      181 ccgagctcat caaggtaaca cttagttttt gttgctctgt ttaaaaatta attttaagga
      241 gaacatgcac atgaagctct acatggaggg aaccgtcgac aaccaccact tcaagtgcac
      301 ctccgaggga gagggaaagc catacgaggg aacccaaacc atgcgtatca aggtcgtcga
      361 gggaggacca ctcccattcg ccttcgacat cctcgccacc tccttcctct acggatcgaa
      421 gaccttcatc aaccacaccc aaggaatccc agacttcttc aagcaatcct tcccaggtaa
      481 gttttcttga gacctttact aactaactag gtctcataga ttttcagagg gattcacctg
      541 ggagcgtgtc accacctacg aggacggagg agtcctcacc gccacccaag acacctccct
      601 ccaagacgga tgcctcatct acaacgtcaa gatccgtgga gtcaacttca cctccaacgg
      661 accagtcatg caaaaggtaa gttatgtaga gacctttact aactaactag gtctcaaaca
      721 ttttcagaag accctcggat gggaggcctt caccgagacg ctctacccag ccgacggagg
      781 actggaggga cgtaacgaca tggccctcaa gctcgtcgga ggatcgcacc tcatcgccaa
      841 cgccaagacc acctaccgtt ccaagaagcc agccaagaac ctcaagatgc cagtctacta
      901 cgtcgactac cgtctggagc gtaaggtaag ttctttagag acctttacta actaactagg
      961 tctcagtaat tttcaggagg ccaacaacga gacgtacgtc gagcaacacg aggtcgccgt
     1021 cgcccgttac tgcgacctcc catccaagct cggacacaag ctcaacggtg gcacaggagg
     1081 aacgggcggt agtggaggca ctggtgccgg caacccagct ttcttgtaca aagttggcat
     1141 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1201 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     1261 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1321 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1381 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1441 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1501 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1561 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1621 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1681 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1741 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1801 cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1861 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1921 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     1981 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2041 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2101 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2161 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2221 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2281 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2821 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2881 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2941 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3001 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3061 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3121 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3181 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     3241 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     3301 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     3361 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     3421 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
     3481 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
