LOCUS       sequence-318050-        3557 bp    DNA     circular SYN 15-APR-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 3557)
  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 3557)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCFJ2301 - [1-2] ENTR - Fluor - ce-mMaple3(1 i...
FEATURES             Location/Qualifiers
     source          1..3557
                     /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"
     CDS             1067..1138
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(1146..1245)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1262..1281)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1263..1281)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1286..1302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1286..1302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1404..1423)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1415..2224
                     /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(1482..1501)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2371..2959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2860..2879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3113..3130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3289..3316)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3408..3494)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3543..3557,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3557,1..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 cgtctccaag gtaacactta
      181 gtttttgttg ctctgtttaa aaattaattt taagggagag gagactatca tgtccgtcat
      241 caagccagac atgaagatca agctccgtat ggagggaaac gtcaacggac acgccttcgt
      301 catcgaggga gagggatccg gaaagccatt cgagggaatc caaaccatcg acctcgaggt
      361 caaggtaagt tttcttgaga cctttactaa ctaactaggt ctcatagatt ttcaggaggg
      421 agccccactc ccattcgcct acgacatcct caccaccgcc ttccactacg gaaaccgtgt
      481 cttcaccaag tacccacgaa agatcccaga ctacttcaag caatccttcc caggtaagtt
      541 atgtagagac cttaatactc aactatggtc tcaaacattt tcagagggat actcctggga
      601 gcgttccatg acctacgagg acggaggaat ctgcaacgcc accaacgaca tcaccatgga
      661 ggaggactcc ttcatcaaca agatccactt caagggaacc aacttcccac caaacggacc
      721 agtcatgcaa aagcgtaccg tcggatggga ggtttccacc gagaaggtaa gttctttaga
      781 gacctaatca catcaatttg gtctcagtaa ttttcagatg tacgtccgtg acggagtcct
      841 caagggtgac gtcaagatga agctcctcct caagggagga tcccactacc gttgcgactt
      901 ccgtaccacc tacaaggtca agcaaaaggc cgtcaagctc ccaaaggccc acttcgtcga
      961 ccaccgtatc gagatcctct cccacgacaa ggactacaac aaggtcaagc tctacgagca
     1021 cgccgtcgcc cgtaactcca ccgactccat ggacgagctc tacaagtccc gtcgtcgtaa
     1081 ggccaaccca accaagctct ccgagaacgc caagaagctc gccaaggagg tcgagaacgg
     1141 atccaaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1201 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1261 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1321 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1381 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1441 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1501 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1561 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1621 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1681 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     1741 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1801 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca
     1861 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1921 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     1981 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2041 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2101 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2161 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2221 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2281 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3241 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     3301 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     3361 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     3421 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     3481 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     3541 ttcccagtca cgacgtt
//
