LOCUS       sequence-318006-        3560 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 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 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: pCFJ2306 - [1-2] ENTR - Fluor - ceGFP(1 intron...
FEATURES             Location/Qualifiers
     source          1..3560
                     /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             1070..1141
                     /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(1149..1248)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1265..1284)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1266..1284)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1289..1305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1289..1305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1407..1426)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1418..2227
                     /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(1485..1504)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2374..2962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2863..2882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3116..3133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3292..3319)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3411..3497)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3546..3560,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3560,1..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 gtccaaggta acacttagtt
      181 tttgttgctc tgtttaaaaa ttaattttaa gggagaggag ctgttcaccg gagtcgtccc
      241 aatcctcgtc gagctcgacg gggacgtcaa cggacacaag ttctccgtgt ccggagaggg
      301 agagggagat gccacctacg gaaagctcac cctcaagttc atctgcacca ccggaaagct
      361 cccagtccca tggccaaccc tcgtcaccac cttctgctac ggagtccaat gcttctcccg
      421 ttacccaggt aagttttctt gagaccttta ctaactaact aggtctcata gattttcaga
      481 ccacatgaag cgtcacgact tcttcaagtc cgccatgcca gagggatacg tccaagagcg
      541 taccatcttc ttcaaggacg acggaaacta caaggtaagt tatgtagaga cctttactaa
      601 ctaactaggt ctcaaacatt ttcagacccg tgccgaggtc aagttcgagg gagacaccct
      661 cgtcaaccgt atcgagctca agggaatcga cttcaaggag gacggtaaca tcctgggaca
      721 caagctggag tacaactaca actcccacaa cgtctacatc atggccgaca agcaaaagaa
      781 cggaatcaag gtcaacttca aggtaagttc tttagagacc tttactaact aactaggtct
      841 cagtaatttt cagatccgtc acaacatcga ggacggatcg gtccaactcg ccgaccacta
      901 ccaacaaaac accccaatcg gtgacggacc agtcctcctc ccagacaacc actacctctc
      961 cacccaatcc gccctctcca aggacccaaa cgagaagcgt gaccacatgg tcctcctcga
     1021 gttcgtcacc gccgccggaa tcacccacgg aatggacgag ctctacaagt cccgtcgtcg
     1081 taaggccaac ccaaccaagc tctccgagaa cgccaagaag ctcgccaagg aggtcgagaa
     1141 cggatccaac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1201 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1261 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1321 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1381 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1441 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1501 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1561 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1621 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1681 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1741 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1801 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc
     1861 tcaggcgcaa tcamgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1921 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     1981 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2041 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2101 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2161 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2221 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2281 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3001 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3061 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3121 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3181 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3241 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     3301 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     3361 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     3421 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     3481 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     3541 gttttcccag tcacgacgtt
//
