LOCUS       sequence-318057-        3536 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 3536)
  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 3536)
  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: pCFJ2300 - [1-2] ENTR - Fluor - ce-Dendra2(1 i...
FEATURES             Location/Qualifiers
     source          1..3536
                     /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"
     CDS             141..161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1046..1117
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(1125..1224)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1241..1260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1242..1260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1265..1281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1265..1281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1383..1402)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1394..2203
                     /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(1461..1480)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2350..2938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2839..2858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3092..3109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3268..3295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3387..3473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3522..3536,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3536,1..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 ctcgggatcc ccaaagaaga agcgtaaggt caacacccca ggtaacactt
      181 agtttttgtt gctctgttta aaaattaatt ttaaggaatc aacctcatca aggaggacat
      241 gcgtgtcaag gtccacatgg agggaaacgt caacggacac gccttcgtca tcgagggaga
      301 gggaaaggga aagccatacg agggaaccca aaccgccaac ctcaccgtca aggtaagttt
      361 tcttgagacc tttactaact aactaggtct catagatttt caggagggag ccccactccc
      421 attctcctac gacatcctca ccaccgccgt ccactacgga aaccgtgtct tcaccaagta
      481 cccagaggac atcccagact acttcaagca atccttccca gagggatact cctgggagcg
      541 taccatgacc ttcgaggaca agggaatctg caccatccgt tccgacatct ccctcgaggg
      601 agactgcttc ttccaaaacg tccgtttcaa ggtaagttat gtagagacct ttactaacta
      661 actaggtctc aaacattttc agggaaccaa cttcccacca aacggaccag tcatgcaaaa
      721 gaagaccctc aagtgggagc catccaccga gaagctccac gtccgtgacg gactcctcgt
      781 cggaaacatc aacatggccc tcctcctcga gggaggagga cactacctct gcgacttcaa
      841 ggtaagttct ttagagacct ttactaacta actaggtctc agtaattttc agaccaccta
      901 caaggccaag aaggtcgtcc aactcccaga cgcccacttc gtcgaccacc gtatcgagat
      961 cctcggaaac gactccgact acaacaaggt caagctctac gagcacgccg tcgcccgtta
     1021 ctccccactc ccatcccaag tctggtcccg tcgtcgtaag gccaacccaa ccaagctctc
     1081 cgagaacgcc aagaagctcg ccaaggaggt cgagaacgga tccaacccag ctttcttgta
     1141 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1201 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     1261 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     1321 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     1381 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     1441 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1501 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1561 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1621 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1681 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1741 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1801 ttgtcctttt aacagcgatc gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa
     1861 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1921 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     1981 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2041 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2101 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2161 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     2221 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     2281 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3001 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3061 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     3181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
     3241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     3301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     3361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     3421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
     3481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgtt
//
