LOCUS       sequence-309273-        6234 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 6234)
  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 6234)
  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: pCFJ2437 - [1-2] ENTR - Fluor -
            ce-tagRFP-T(PA...
FEATURES             Location/Qualifiers
     source          1..6234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(143..162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(798..1607)
                     /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     1521..1540
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1599..1618
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1720..1736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1720..1736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1741..1760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1741..1759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1777..1876
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5567..5666)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5682..5699)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5682..5698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5691..5713)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      5762..5848
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5940..5967
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6126..6143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
       61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat
      721 tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
      781 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
      841 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
      901 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
      961 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1021 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     1081 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1141 ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa
     1201 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1261 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg
     1321 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1381 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1441 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     1501 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1561 ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac accccttgta
     1621 ttactgttta tgtaagcaga cagttttatt gttcatgatg atattatttt atcttgtgca
     1681 atgtaacatc agagattttg agacacgggc cagagctgcc aggaaacagc tatgaccatg
     1741 taatacgact cactataggg gatatcagct ggatggcaaa taatgatttt attttgactg
     1801 atagtgacct gttcgttgca acaaattgat aagcaatgct ttcttataat gccaactttg
     1861 tacaagaaag ctgggttgcc ggcaccagtg cctccactac cgcccgttcc tcctgtgcca
     1921 cccttgtaga gctcgtccat tccgttgagc ttgtgtccga gcttggatgg gaggtcgcag
     1981 taacgggcga cggcgacctc gtgttgctcg acgtacgtct ccttgtcggc ctccttgata
     2041 cgctccagac ggtggtcgac gtagtagact cctggcatct tgaggttctg aaaattactt
     2101 tggggatttt tagtcggaaa aatggatttt ttggggaaaa atcgggattt tctagtgaaa
     2161 aatcacgaaa aactggaatt ttgtgttaaa atcagtgatt tttactctaa aaatcactga
     2221 aaaactgatt ttttggttga aaatccaatt tttttcgatt aaaaatcaaa tttttggctc
     2281 aaaaatcgga attttcagtc taaactcatt tttaaacggg gtttttttgg aaaatttcga
     2341 gttttttggc tgaaaatcca gaatttcagg gatttttagc tgaaaattga ctgaaaaacg
     2401 ataattttca gcggaaaaat aaaatttttg gtctaaaaac aacttgattt tctagtgaaa
     2461 atccagtttt ttttcagtga aaaagtggaa aaatctatat tttttcaggc aaaattagtg
     2521 atttttgatc gaaaattgat ttttagtcta aaaaaaagtg tttttttggc aaaaagttca
     2581 ggttttcacg taaaaaacta aaaacctcaa atttttggct taaaaatcgg agtttttgtc
     2641 gaaaatgctg tgtttttctt gtgaaatagc actaaaaact gaatttttac ccccgtaaat
     2701 ccacagtttt tcgagtaaaa atagaaaatt cagggatttt ttgtgtgtga aaaattgaaa
     2761 tttggccatt tttcagaatt tttggccttt ttaggcgatt ttcggccttt attaggtaat
     2821 ttttggcctt ttttcggtga tttttattaa tttttagcgg gaatttcctc aaaatttgag
     2881 attttcggcg attttcagat aatttttggc cgtttttcgg tgatttttag gtcatttttc
     2941 agaattttcg gccaattttg aggattcttt ggccattttt tgtgaattcc atcaattttt
     3001 cactcaaaaa cgagcaaaaa aaagaactta ccttggctgg cttcttggaa cggtaggtgg
     3061 tcttgaagtt gcagatgagg tgtcctcctc cgacgagctt gagggccatg tcggtacgtc
     3121 cctccagtcc tccgtcggct gggtagagca tctcggtgtt ggcctcccat ccgagggtct
     3181 tcttttgcat gactggtccg ttggatggga agttgactcc acggatctga aaatgttaat
     3241 cgataaattt ggagcttttt tcgtcttttt ccggtgaaaa gtggggaatt ttgagctttt
     3301 tttgaagatt ttcagcctaa aatcgtcatt tttcggtgat ttttgcacaa ttttcagcag
     3361 caaaaattag ggagaattta gacgttttga agccaatttt tgtaaaaaaa atcgcatttt
     3421 tctgattgat tttttttttg tcatttttgg agaaaaaaaa gggcaatctg agtattttta
     3481 gccaaaaaat ttgaatttta atgtttttaa agcaatttta gccgaaaata ggcagatttt
     3541 cagctaaaaa ttgaaaattc agtctttttc gacgatttta gctcaatttc cggtgaaaat
     3601 tgggtaattt tgagcatttt tcaagttttt ttaagctcaa aattgccaat ttcggcagtt
     3661 tttttagcgg tttttcggcg atttttgctc aatttttcaa aaaaatttca atttttccac
     3721 gaaaaacgcc agaaaagtcc gattttggag atttttggcc aatttttcgg cttttgaact
     3781 ttttcagaca aaaatcggtg tttttcatat atttttcgtc aaatttccgc caaaaattga
     3841 aaatttccga attttctgct ggaaaatcca attttccacc attttcagtg tgaaaaaagc
     3901 tcaaaaagtc tgtttttgag caaaaaagtc catttttcac aattttcggt tcaaaatcta
     3961 ccaattttcg ccgaaaaacg tcgattttgg caggaaattg cgattctgaa caattttctg
     4021 gctgaaattt cagcaaaaaa ggcatttttc gtcgatttct gcaatatttt cagctttttt
     4081 cggtgaattt tcagcaaaaa aaccaatttt cagcattttc agtgcaaaaa aagctgaaaa
     4141 attcagtttt tgagcaaaaa agtccatttt ttcgccgaga aatcgcgtac ataacttacc
     4201 ttgacgttgt agatgaggca tccgtcttgg agggaggtgt cttgggtggc ggtgaggact
     4261 cctccgtcct cgtaggtggt gacacgctcc caggtgaatc cctctgggaa ggattgcttg
     4321 aagaagtctg ggattcctga aaatctaatt tttggtaaaa aatcgatttt ttttttcgtc
     4381 tttcgtccca cttttgagct aaaaatccca atttttcagt tgaaaattgt attttttagc
     4441 taaaaatctc acttttcagc taaaaacatc tcacttttca tcttaaaatc ccactttttt
     4501 acttaaaaat tccaattttt cagctaaaca tttcaatttt tcaactaaaa atctcacttt
     4561 tcagctaaaa atccaatttt taaagctgaa aatccaattt ttaaagctaa aaatcccaat
     4621 ttttcagttg aaaattgtat tttttagctg aaaatctcac ttttcagcta aaaaatctca
     4681 cttttcatct taaaatccca ctttttaact aaaaaatttc aattttcagc taaaaatccg
     4741 attttttagc taaaaatcca atttttaaag ctaaaaatca cgatctttag ttgaaaattg
     4801 tattttttag ctaaaaatat tattttttaa ctaaaaatct cacttttctg ttaaaaaatc
     4861 tcacttttta actaaaaaat ttcaattttc agctaaaaat ccgatttttc agctaaaaat
     4921 ccaattttta aagctaaaaa tcacgatttt tagttgaaaa tggaattttt tagctaaaaa
     4981 ttcaaatttt tccgcttaaa atttgattta tcagctaaaa atacaatttt ttctgcttaa
     5041 aattccaatc tttcagctaa aaatcatact ttttagctag aaatctcgat ttttcaacta
     5101 aaaatcccac tctttagcta aaaatttaac ttttcagcta aaaattgaaa tttttagttg
     5161 aaaatttcaa tttttcatct taaaatccaa tttttcagct aaaaatccca attctcagaa
     5221 acaagtaaat aaaacagaaa acttaccttg ggtgtggttg atgaaggtac gcgatccgta
     5281 catgaaggag gtggcgagga tgtcgaaggc gaatgggagt ggtcctccct cgacgacctt
     5341 gatacgcatg gtttgggttc cctcgtatgg ctttccctct ccctcggagg tgcacttgaa
     5401 gtggtggttg ttgacggttc cctccatgta gagcttcatg tgcatgttct ccttgatgag
     5461 ctcctctccc ttaaaattaa tttttaaaca gagcaacaaa aactaagtgt taccttggaa
     5521 ctgcttccgc cggtacctcc actgccaccg ctagtactgc cggccgagcc tgcttttttg
     5581 tacaaagttg gcattataaa aaagcattgc tcatcaattt gttgcaacga acaggtcact
     5641 atcagtcaaa ataaaatcat tatttggggc ccgagcttaa gactggccgt cgttttacaa
     5701 cgtcgtgact gggaaaacat ccatgctagc gttaacgcga gagtagggaa ctgccaggca
     5761 tcaaataaaa cgaaaggctc agtcggaaga ctgggccttt cgttttatct gttgtttgtc
     5821 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgtgaagca
     5881 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaactaagca
     5941 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttcctg gctagcggta
     6001 cgcgtattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6061 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaag
//
