LOCUS       sequence-318055-        6216 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 6216)
  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 6216)
  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: pCFJ2443 - [1-2] ENTR - Fluor -
            ce-mMaple3(PAT...
FEATURES             Location/Qualifiers
     source          1..6216
                     /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(3805..3904)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3921..3940)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3922..3940)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3945..3961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3945..3961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4063..4082)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4074..4883
                     /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(4141..4160)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5030..5618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5519..5538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5772..5789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5948..5975)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6067..6153)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6202..6216,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6216,1..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 tctgttttat ttacttgttt ctgagaattg
      421 ggatttttag ctgaaaaatt ggattttaag atgaaaaatt gaaattttca actaaaaatt
      481 tcaattttta gctgaaaagt taaattttta gctaaagagt gggattttta gttgaaaaat
      541 cgagatttct agctaaaaag tatgattttt agctgaaaga ttggaatttt aagcagaaaa
      601 aattgtattt ttagctgata aatcaaattt taagcggaaa aatttgaatt tttagctaaa
      661 aaattccatt ttcaactaaa aatcgtgatt tttagcttta aaaattggat ttttagctga
      721 aaaatcggat ttttagctga aaattgaaat tttttagtta aaaagtgaga ttttttaaca
      781 gaaaagtgag atttttagtt aaaaaataat atttttagct aaaaaataca attttcaact
      841 aaagatcgtg atttttagct ttaaaaattg gatttttagc taaaaaatcg gatttttagc
      901 tgaaaattga aattttttag ttaaaaagtg ggattttaag atgaaaagtg agatttttta
      961 gctgaaaagt gagattttca gctaaaaaat acaattttca actgaaaaat tgggattttt
     1021 agctttaaaa attggatttt cagctttaaa aattggattt ttagctgaaa agtgagattt
     1081 ttagttgaaa aattgaaatg tttagctgaa aaattggaat ttttaagtaa aaaagtggga
     1141 ttttaagatg aaaagtgaga tgtttttagc tgaaaagtga gatttttagc taaaaaatac
     1201 aattttcaac tgaaaaattg ggatttttag ctcaaaagtg ggacgaaaga cgaaaaaaaa
     1261 aatcgatttt ttaccaaaaa ttagattttc aggagggagc cccactccca ttcgcctacg
     1321 acatcctcac caccgccttc cactacggaa accgtgtctt caccaagtac ccacgaaaga
     1381 tcccagacta cttcaagcaa tccttcccag gtaagttatg tacgcgattt ctcggcgaaa
     1441 aaatggactt ttttgctcaa aaactgaatt tttcagcttt ttttgcactg aaaatgctga
     1501 aaattggttt ttttgctgaa aattcaccga aaaaagctga aaatattgca gaaatcgacg
     1561 aaaaatgcct tttttgctga aatttcagcc agaaaattgt tcagaatcgc aatttcctgc
     1621 caaaatcgac gtttttcggc gaaaattggt agattttgaa ccgaaaattg tgaaaaatgg
     1681 acttttttgc tcaaaaacag actttttgag cttttttcac actgaaaatg gtggaaaatt
     1741 ggattttcca gcagaaaatt cggaaatttt caatttttgg cggaaatttg acgaaaaata
     1801 tatgaaaaac accgattttt gtctgaaaaa gttcaaaagc cgaaaaattg gccaaaaatc
     1861 tccaaaatcg gacttttctg gcgtttttcg tggaaaaatt gaaatttttt tgaaaaattg
     1921 agcaaaaatc gccgaaaaac cgctaaaaaa actgccgaaa ttggcaattt tgagcttaaa
     1981 aaaacttgaa aaatgctcaa aattacccaa ttttcaccgg aaattgagct aaaatcgtcg
     2041 aaaaagactg aattttcaat ttttagctga aaatctgcct attttcggct aaaattgctt
     2101 taaaaacatt aaaattcaaa ttttttggct aaaaatactc agattgccct tttttttctc
     2161 caaaaatgac aaaaaaaaaa tcaatcagaa aaatgcgatt ttttttacaa aaattggctt
     2221 caaaacgtct aaattctccc taatttttgc tgctgaaaat tgtgcaaaaa tcaccgaaaa
     2281 atgacgattt taggctgaaa atcttcaaaa aaagctcaaa attccccact tttcaccgga
     2341 aaaagacgaa aaaagctcca aatttatcga ttaacatttt cagagggata ctcctgggag
     2401 cgttccatga cctacgagga cggaggaatc tgcaacgcca ccaacgacat caccatggag
     2461 gaggactcct tcatcaacaa gatccacttc aagggaacca acttcccacc aaacggacca
     2521 gtcatgcaaa agcgtaccgt cggatgggag gtttccaccg agaaggtaag ttcttttttt
     2581 tgctcgtttt tgagtgaaaa attgatggaa ttcacaaaaa atggccaaag aatcctcaaa
     2641 attggccgaa aattctgaaa aatgacctaa aaatcaccga aaaacggcca aaaattatct
     2701 gaaaatcgcc gaaaatctca aattttgagg aaattcccgc taaaaattaa taaaaatcac
     2761 cgaaaaaagg ccaaaaatta cctaataaag gccgaaaatc gcctaaaaag gccaaaaatt
     2821 ctgaaaaatg gccaaatttc aatttttcac acacaaaaaa tccctgaatt ttctattttt
     2881 actcgaaaaa ctgtggattt acgggggtaa aaattcagtt tttagtgcta tttcacaaga
     2941 aaaacacagc attttcgaca aaaactccga tttttaagcc aaaaatttga ggtttttagt
     3001 tttttacgtg aaaacctgaa ctttttgcca aaaaaacact tttttttaga ctaaaaatca
     3061 attttcgatc aaaaatcact aattttgcct gaaaaaatat agatttttcc actttttcac
     3121 tgaaaaaaaa ctggattttc actagaaaat caagttgttt ttagaccaaa aattttattt
     3181 ttccgctgaa aattatcgtt tttcagtcaa ttttcagcta aaaatccctg aaattctgga
     3241 ttttcagcca aaaaactcga aattttccaa aaaaaccccg tttaaaaatg agtttagact
     3301 gaaaattccg atttttgagc caaaaatttg atttttaatc gaaaaaaatt ggattttcaa
     3361 ccaaaaaatc agtttttcag tgatttttag agtaaaaatc actgatttta acacaaaatt
     3421 ccagtttttc gtgatttttc actagaaaat cccgattttt ccccaaaaaa tccatttttc
     3481 cgactaaaaa tccccaaagt aattttcaga tgtacgtccg tgacggagtc ctcaagggtg
     3541 acgtcaagat gaagctcctc ctcaagggag gatcccacta ccgttgcgac ttccgtacca
     3601 cctacaaggt caagcaaaag gccgtcaagc tcccaaaggc ccacttcgtc gaccaccgta
     3661 tcgagatcct ctcccacgac aaggactaca acaaggtcaa gctctacgag cacgccgtcg
     3721 cccgtaactc caccgactcc atggacgagc tctacaaggg tggcacagga ggaacgggcg
     3781 gtagtggagg cactggtgcc ggcaacccag ctttcttgta caaagttggc attataagaa
     3841 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     3901 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     3961 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4021 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     4081 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     4141 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     4201 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     4261 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     4321 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     4381 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     4441 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     4501 gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     4561 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     4621 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     4681 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     4741 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     4801 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     4861 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     4921 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     4981 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5041 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5101 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5161 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5221 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5281 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5341 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5401 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5461 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5521 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5581 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5641 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5701 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5761 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5821 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5881 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     5941 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     6001 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     6061 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     6121 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt
     6181 taacgctagc atggatgttt tcccagtcac gacgtt
//
