LOCUS       sequence-318036-        3575 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 3575)
  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 3575)
  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: pCFJ2309 - [1-2] ENTR - Fluor - mCardinal(1 in...
FEATURES             Location/Qualifiers
     source          1..3575
                     /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             1085..1156
                     /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(1164..1263)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1280..1299)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1281..1299)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1304..1320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1304..1320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1422..1441)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1433..2242
                     /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(1500..1519)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2389..2977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2878..2897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3131..3148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3307..3334)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3426..3512)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3561..3575,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3575,1..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 tatattgcca actttgtaca
      121 aaaaagcagg ctcgggatcc ccaaagaaga agcgtaaggt gtccaaggta acacttagtt
      181 tttgttgctc tgtttaaaaa ttaattttaa gggagaggag ctcatcaagg agaacatgca
      241 catgaagctc tacatggagg gaaccgtcaa caaccaccac ttcaagtgca ccaccgaggg
      301 agagggaaag ccatacgagg gaacccaaac ccaacgtatc aaggtcgtcg agggaggacc
      361 actcccattc gccttcgaca tcctcgccac ctgcttcatg tacggatcga aggtaagttt
      421 tcttgagacc tttactaact aactaggtct catagatttt cagaccttca tcaaccacac
      481 ccaaggaatc ccagacttct tcaagcaatc cttcccagag ggattcacct gggagcgtgt
      541 caccacctac gaggacggag gagtcctcac cgtcacccaa gacacctccc tccaaggtaa
      601 gttatgtaga gacctttact aactaactag gtctcaaaca ttttcagacg gatgcctcat
      661 ctacaacgtc aagctccgtg gagtcaactt cccatccaac ggaccagtca tgcaaaagaa
      721 gaccctcgga tgggaggcca ccaccgagac gctctaccca gccgacggag gactcgaggg
      781 acgttgcgac atggccctca agctcgtcgg aggaggacac ctccactgca acctcaaggt
      841 aagttcttta gagaccttta ctaactaact aggtctcagt aattttcaga ccacctaccg
      901 ttccaagaag ccagccaaga acctcaagat gccaggagtc tacttcgtcg accgtcgtct
      961 cgagcgtatc aaggaggccg acaacgagac gtacgtcgag caacacgagg tcgccgtcgc
     1021 ccgttactgc gacctcccat ccaagctcgg acacaagctc aacggaatgg acgagctcta
     1081 caagtcccgt cgtcgtaagg ccaacccaac caagctctcc gagaacgcca agaagctcgc
     1141 caaggaggtc gagaacggat ccaacccagc tttcttgtac aaagttggca ttataagaaa
     1201 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1261 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1321 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1381 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     1441 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     1501 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     1561 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1621 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1681 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     1741 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1801 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     1861 cgtatttcgc ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     1921 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     1981 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2041 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2101 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2161 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2221 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     2281 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     2341 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3001 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3061 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3121 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3181 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3241 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     3301 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3361 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3421 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3481 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     3541 aacgctagca tggatgtttt cccagtcacg acgtt
//
