LOCUS       sequence-317995-        3740 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 3740)
  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 3740)
  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: pCFJ1972 - [1-2] ENTR - Flour - TagBFP2(NLS(2)...
FEATURES             Location/Qualifiers
     source          1..3740
                     /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"
     CDS             141..161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1328..1427)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1444..1463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1445..1463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1468..1484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1468..1484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1586..1605)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1597..2406
                     /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(1664..1683)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2553..3142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3043..3062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3296..3313
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3472..3499)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3591..3677)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3726..3740,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3740,1..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 ctcgggatcc ccaaagaaga agcgtaaggt aagttttgaa atattcatat
      181 ttttaacttg aaaactaata tttaaatttt ttaggtcgcc ggcggatcat cctccacatc
      241 atcctccaca tcatcctcat ccgagctcat caaggtaaca cttagttttt gttgctctgt
      301 ttaaaaatta attttaagga gaacatgcac atgaagctct acatggaggg aaccgtcgac
      361 aaccaccact tcaagtgcac ctccgaggga gagggaaagc catacgaggg aacccaaacc
      421 atgcgtatca aggtcgtcga gggaggacca ctcccattcg ccttcgacat cctcgccacc
      481 tccttcctct acggatcgaa gaccttcatc aaccacaccc aaggaatccc agacttcttc
      541 aagcaatcct tcccaggtaa gttttcttga gacctttact aactaactag gtctcataga
      601 ttttcagagg gattcacctg ggagcgtgtc accacctacg aggacggagg agtcctcacc
      661 gccacccaag acacctccct ccaagacgga tgcctcatct acaacgtcaa gatccgtgga
      721 gtcaacttca cctccaacgg accagtcatg caaaaggtaa gttatgtaga gacctttact
      781 aactaactag gtctcaaaca ttttcagaag accctcggat gggaggcctt caccgagacg
      841 ctctacccag ccgacggagg actggaggga cgtaacgaca tggccctcaa gctcgtcgga
      901 ggatcgcacc tcatcgccaa cgccaagacc acctaccgtt ccaagaagcc agccaagaac
      961 ctcaagatgc cagtctacta cgtcgactac cgtctggagc gtaaggtaag ttctttagag
     1021 acctttacta actaactagg tctcagtaat tttcaggagg ccaacaacga gacgtacgtc
     1081 gagcaacacg aggtcgccgt cgcccgttac tgcgacctcc catccaagct cggacacaag
     1141 ctcaactcgg gaacttctac gacttcggta acgtcatccg gtacttcccc agccggctcc
     1201 cgtcgtcgta aggtaagttc ataagagacc tttactaact aactaggtct cagaattttt
     1261 caggccaacc caaccaagct ctccgagaac gccaagaagc tcgccaagga ggtcgagaac
     1321 ggatccaacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     1381 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     1441 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1501 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1561 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1621 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1681 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1741 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1801 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1861 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1921 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1981 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgcctcgct
     2041 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2101 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2161 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2221 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2281 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2341 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2401 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     2461 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2581 tgctgcttgc aaacaaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3181 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3241 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3301 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3361 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3421 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     3481 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     3541 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     3601 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     3661 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     3721 gttttcccag tcacgacgtt
//
