LOCUS       sequence-326622-        3560 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 3560)
  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 3560)
  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: pCFJ2240 - [1-2] ENTR - Fluor - ce-tagRFP-T (1...
FEATURES             Location/Qualifiers
     source          1..3560
                     /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(1685..1784)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1801..1820)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1802..1820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1825..1841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1825..1841)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1943..1962)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1954..2763
                     /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(2021..2040)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2910..3498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3399..3418
                     /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 gcagttccaa
      721 ggtaacactt agtttttgtt gctctgttta aaaattaatt ttaagggaga ggagctcatc
      781 aaggagaaca tgcacatgaa gctctacatg gagggaaccg tcaacaacca ccacttcaag
      841 tgcacctccg agggagaggg aaagccatac gagggaaccc aaaccatgcg tatcaaggtc
      901 gtcgagggag gaccactccc attcgccttc gacatcctcg ccacctcctt catgtacgga
      961 tcgcgtacct tcatcaacca cacccaaggt aagttttctt gagaccttta ctaactaact
     1021 aggtctcata gattttcagg aatcccagac ttcttcaagc aatccttccc agagggattc
     1081 acctgggagc gtgtcaccac ctacgaggac ggaggagtcc tcaccgccac ccaagacacc
     1141 tccctccaag acggatgcct catctacaac gtcaaggtaa gttatgtaga gacctttact
     1201 aactaactag gtctcaaaca ttttcagatc cgtggagtca acttcccatc caacggacca
     1261 gtcatgcaaa agaagaccct cggatgggag gccaacaccg agatgctcta cccagccgac
     1321 ggaggactgg agggacgtac cgacatggcc ctcaagctcg tcggaggagg acacctcatc
     1381 tgcaacttca agaccaccta ccgttccaag aagccagcca aggtaagttc tttagagacc
     1441 tttactaact aactaggtct cagtaatttt cagaacctca agatgccagg agtctactac
     1501 gtcgaccacc gtctggagcg tatcaaggag gccgacaagg agacgtacgt cgagcaacac
     1561 gaggtcgccg tcgcccgtta ctgcgacctc ccatccaagc tcggacacaa gctcaacgga
     1621 atggacgagc tctacaaggg tggcacagga ggaacgggcg gtagtggagg cactggtgcc
     1681 ggcaacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     1741 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     1801 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1861 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1921 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     1981 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2041 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2101 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2161 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2221 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2281 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2341 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg cctcgctcag
     2401 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2461 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2521 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2581 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2641 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2701 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2761 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     2821 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     2881 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2941 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3001 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3061 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3121 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3181 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3241 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3301 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3361 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3421 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3481 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3541 tggccttttg ctcacatgtt
//
