LOCUS       sequence-318048-        3542 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 3542)
  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 3542)
  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: pCFJ2210 - [1-2] ENTR - Fluor - mMaple3(1 intr...
FEATURES             Location/Qualifiers
     source          1..3542
                     /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"
     protein_bind    complement(1131..1230)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1247..1266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1248..1266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1271..1287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1271..1287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1389..1408)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1400..2209
                     /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(1467..1486)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2356..2944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2845..2864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3098..3115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3274..3301)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3393..3479)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3528..3542,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3542,1..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 tataatgcca actttgtaca
      121 aaaaagcagg ctcggccggc agtactagcg gtggcagtgg aggtaccggc ggaagcagtg
      181 tctccaaggt aacacttagt ttttgttgct ctgtttaaaa attaatttta agggagagga
      241 gactatcatg tccgtcatca agccagacat gaagatcaag ctccgtatgg agggaaacgt
      301 caacggacac gccttcgtca tcgagggaga gggatccgga aagccattcg agggaatcca
      361 aaccatcgac ctcgaggtca aggtaagttt tcttgagacc tttactaact aactaggtct
      421 catagatttt caggagggag ccccactccc attcgcctac gacatcctca ccaccgcctt
      481 ccactacgga aaccgtgtct tcaccaagta cccacgaaag atcccagact acttcaagca
      541 atccttccca ggtaagttat gtagagacct taatactcaa ctatggtctc aaacattttc
      601 agagggatac tcctgggagc gttccatgac ctacgaggac ggaggaatct gcaacgccac
      661 caacgacatc accatggagg aggactcctt catcaacaag atccacttca agggaaccaa
      721 cttcccacca aacggaccag tcatgcaaaa gcgtaccgtc ggatgggagg tttccaccga
      781 gaaggtaagt tctttagaga cctaatcaca tcaatttggt ctcagtaatt ttcagatgta
      841 cgtccgtgac ggagtcctca agggtgacgt caagatgaag ctcctcctca agggaggatc
      901 ccactaccgt tgcgacttcc gtaccaccta caaggtcaag caaaaggccg tcaagctccc
      961 aaaggcccac ttcgtcgacc accgtatcga gatcctctcc cacgacaagg actacaacaa
     1021 ggtcaagctc tacgagcacg ccgtcgcccg taactccacc gactccatgg acgagctcta
     1081 caagggtggc acaggaggaa cgggcggtag tggaggcact ggtgccggca acccagcttt
     1141 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     1201 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1261 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1321 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1381 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1441 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1501 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1561 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1621 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1681 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1741 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1801 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat
     1861 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1921 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1981 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2041 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2101 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2161 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2221 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2281 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3061 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     3241 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     3301 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3361 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3421 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     3481 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
     3541 tt
//
