LOCUS       sequence-318032-        4655 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 4655)
  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 4655)
  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: pCFJ1410 - [1-2] ENTR - Fluor - mCherry(PATCs,...
FEATURES             Location/Qualifiers
     source          1..4655
                     /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             2171..2242
                     /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(2244..2343)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2360..2379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2361..2379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2384..2400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2384..2400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2502..2521)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2513..3322
                     /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(2580..2599)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3469..4057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3958..3977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4211..4228
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4387..4414)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4506..4592)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4641..4655,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4655,1..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 gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctcgccaaag gtaagtttat ctaaaaagtt tttcattcaa aatgtgtaaa
      181 aattcattta aaataaccca aaaatcatta atcctcgata ttttcagaag aagcgtaagg
      241 tgtccaaggt gtccaaggga gaggaggata acatggctat catcaaggta ttttcctgca
      301 tttttcaact gggaaaatga aagaaaatcg ataatttcag gagttcatga gattcaaggt
      361 ccacatggag ggatccgtca acggacacga gttcgagatc gagggagagg gagagggaag
      421 accatacgag ggaacccaga ccgctaagct taaggtcacc aaggtaagat atgggaaaga
      481 aggaaaaaac cgagatttta cttgaaaaat tgaatttttc gcgggatttt caccaaaaat
      541 tgttgaatat tcattatttc acgctgtaaa acaaaaataa aaataatcaa aaactacgtt
      601 gaaatcgcgt ttttaagcga attttcttca gaattgccag attttaaccc caaattttgc
      661 agtttttaaa taaaatttca ccttttcggc tcaaattgta gattttcctg aaaatttagt
      721 acaaaaaaca atttcctcgt aaatttttca aattttcagg gaggaccact tccattcgct
      781 tgggatatcc tttccccaca gttcatgtac ggatccaagg cttacgtcaa gcacccagct
      841 gatatcccag attaccttaa gctttccttc ccagagggat tcaagtggga gagagtcatg
      901 aacttcgagg atggaggagt cgtcaccgtc acccaggatt cctcccttca ggatggagag
      961 ttcatctaca aggtaagtta tgtctacctg cctgcctacc gcctaaattt tttgtgaagt
     1021 ttcttcaaaa aatccagaaa aaaaaacaat tttcatacga ttttttccct taaaattgtg
     1081 aattttcatg ctttttagcc ccaaaagtca ttatttgaga aaaaatttca tacaaaaaag
     1141 ttttgagaaa tacacaattt tttaaatgta attttcaaat tttcaatttt caactagaaa
     1201 attcacaaaa cttgtaaatt ttggaccaaa acatttatac aattactttt tttgaatcta
     1261 ataactacaa taactacaaa tttaacattt tcaggtcaag cttagaggaa ccaacttccc
     1321 atccgatgga ccagtcatgc agaagaagac catgggatgg gaggcttcct ccgagagaat
     1381 gtacccagag gatggagctc ttaagggaga gatcaagcag agacttaagc ttaaggatgg
     1441 aggacactac gatgctgagg tcaagaccac ctacaaggta agttcttttt ttgaaaagtg
     1501 cagtttgtag tctaattttc atttattttc tttttaaaaa acgcatcaat ttttaaatat
     1561 tttgggacaa aatccgaaaa ctgtactaat ttgtagtttg taaaattaaa aaaaaaacgc
     1621 aaaaaaatgt tttcaaaaat gcttagaata aaaataaaat ttctaaaaaa ttgaacaaaa
     1681 taaacattta aaaattcaaa aagtttgaaa aaatgcaaag ttttttaatg caaaaaatat
     1741 ttaatgttta aacaaattta aaaaatgttg taagaatatg tttagagaca gtttttaaat
     1801 gttaaattgc aaaaaaaaaa agcaaaaaaa aatttgaaaa atgtaacaga aaattttaat
     1861 taaaaaagtc gtgttttttt caaacctgca attctttgta attttattta taaagagaaa
     1921 caaatcaaat tgtatattac gaaaaacgat actaaaattc gaaaatttgc ggttttttgc
     1981 gtaaaaaata cggttccgta attttcaggc taagaagcca gtccagcttc caggagctta
     2041 caacgtcaac atcaagcttg atatcacctc ccacaacgag gattacacca tcgtcgagca
     2101 gtacgagaga gctgagggaa gacactccac cggaggaatg gatgagcttt acaagggatc
     2161 aggtagtggc tcccgtcgtc gtaaggccaa cccaaccaag ctctccgaga acgccaagaa
     2221 gctcgccaag gaggtcgaga acaacccagc tttcttgtac aaagttggca ttataagaaa
     2281 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2341 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2401 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2461 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2521 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2581 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2641 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2701 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2761 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2821 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2881 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2941 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3001 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3061 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3121 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3181 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3241 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3301 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3361 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3421 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4081 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4141 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4201 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4261 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4321 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     4381 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     4441 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     4501 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     4561 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     4621 aacgctagca tggatgtttt cccagtcacg acgtt
//
