LOCUS       sequence-309277-        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     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             677..697
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1621..1692
                     /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(1700..1799)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1816..1835)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1817..1835)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1840..1856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1840..1856)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1958..1977)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1969..2778
                     /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(2036..2055)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2925..3513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3414..3433
                     /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 agcaggctcg ggatccccaa agaagaagcg taaggtgtcc aaggtaacac ttagtttttg
      721 ttgctctgtt taaaaattaa ttttaaggga gaggagctca tcaaggagaa catgcacatg
      781 aagctctaca tggagggaac cgtcaacaac caccacttca agtgcaccac cgagggagag
      841 ggaaagccat acgagggaac ccaaacccaa cgtatcaagg tcgtcgaggg aggaccactc
      901 ccattcgcct tcgacatcct cgccacctgc ttcatgtacg gatcgaaggt aagttttctt
      961 gagaccttta ctaactaact aggtctcata gattttcaga ccttcatcaa ccacacccaa
     1021 ggaatcccag acttcttcaa gcaatccttc ccagagggat tcacctggga gcgtgtcacc
     1081 acctacgagg acggaggagt cctcaccgtc acccaagaca cctccctcca aggtaagtta
     1141 tgtagagacc tttactaact aactaggtct caaacatttt cagacggatg cctcatctac
     1201 aacgtcaagc tccgtggagt caacttccca tccaacggac cagtcatgca aaagaagacc
     1261 ctcggatggg aggccaccac cgagacgctc tacccagccg acggaggact cgagggacgt
     1321 tgcgacatgg ccctcaagct cgtcggagga ggacacctcc actgcaacct caaggtaagt
     1381 tctttagaga cctttactaa ctaactaggt ctcagtaatt ttcagaccac ctaccgttcc
     1441 aagaagccag ccaagaacct caagatgcca ggagtctact tcgtcgaccg tcgtctcgag
     1501 cgtatcaagg aggccgacaa cgagacgtac gtcgagcaac acgaggtcgc cgtcgcccgt
     1561 tactgcgacc tcccatccaa gctcggacac aagctcaacg gaatggacga gctctacaag
     1621 tcccgtcgtc gtaaggccaa cccaaccaag ctctccgaga acgccaagaa gctcgccaag
     1681 gaggtcgaga acggatccaa cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1741 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1801 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     1861 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaat aatatcatca
     1921 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1981 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     2041 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     2101 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     2161 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     2221 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     2281 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     2341 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2401 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2461 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     2521 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2581 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2641 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2701 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2761 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     2821 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     2881 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2941 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3001 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3061 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3121 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3181 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3241 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3301 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3361 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3421 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3481 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3541 ttcctggcct tttgctggcc ttttgctcac atgtt
//
