LOCUS       sequence-318863-        3773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-fragment Gateway compatible protein tag for expression in C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3773)
  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 3773)
  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: pCFJ2059 - [1-2] ENTR - Tag - Halo (intron(1),...
FEATURES             Location/Qualifiers
     source          1..3773
                     /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"
     protein_bind    complement(1898..1997)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2014..2033)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2015..2033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2038..2054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2038..2054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2156..2175)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2167..2976
                     /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(2234..2253)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3123..3711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3612..3631
                     /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 gccggcagta ctagcggtgg cagtggaggt accggcggaa gcagtgccga
      721 ggtaacactt agtttttgtt gctctgttta aaaattaatt ttaagatcgg aaccggattc
      781 ccattcgacc cacactacgt cgaggtcctc ggagagcgta tgcactacgt cgacgtcgga
      841 ccacgtgacg gaaccccagt cctcttcctc cacggaaacc caacctcctc ctacgtctgg
      901 cgtaacatca tcccacacgt cgccccaacc caccgttgca tcgccccagg taagttttct
      961 tgagaccttt actaactaac taggtctcat agattttcag acctcatcgg aatgggaaag
     1021 tccgacaagc cagacctcgg atacttcttc gacgaccacg tccgtttcat ggacgccttc
     1081 atcgaggccc tcggactcga ggaggtcgtc ctcgtcatcc acgactgggg atcggctctc
     1141 ggattccact gggccaagcg taacccagag cgtgtcaagg gaatcgcctt catggagttc
     1201 atccgtccaa tcccaacctg ggacgagtgg ccaggtaagt tatgtagaga cctttactaa
     1261 ctaactaggt ctcaaacatt ttcagagttc gcccgtgaga cattccaagc cttccgtacc
     1321 accgacgtcg gacgtaagct catcatcgac caaaacgtct tcatcgaggg aaccctccca
     1381 atgggagtcg tccgtccact caccgaggtc gagatggacc actaccgtga gccattcctc
     1441 aacccagtcg accgtgagcc actctggcgt ttcccaaacg agctcccaat cgccggagag
     1501 ccagccaaca tcgtcgccct cgtcgaggag tacatggact ggctccacca atccccagtc
     1561 ccaaagctcc tcttctgggg aaccccaggt aagttcttta gagaccttta ctaactaact
     1621 aggtctcagc aattttcagg agtcctcatc ccaccagccg aggccgcccg tctcgccaag
     1681 tccctcccaa actgcaaggc cgtcgacatc ggaccaggac tcaacctcct ccaagaggac
     1741 aacccaggta agttcataag agacctttac taactaacta ggtctcagaa tttttcagac
     1801 ctcatcggtt ccgagatcgc ccgttggctc tccaccctcg agatctccgg aggtggcaca
     1861 ggaggaacgg gcggtagtgg aggcactggt gccggcaacc cagctttctt gtacaaagtt
     1921 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1981 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     2041 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     2101 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2161 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     2221 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2281 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2341 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2401 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2461 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2521 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2581 tttaacagcg atcgcgtatt tcgcctcgct caggcgcaat cacgaatgaa taacggtttg
     2641 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2701 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2761 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2821 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2881 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2941 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     3001 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     3061 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3241 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
