LOCUS       sequence-317999-        5206 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 5206)
  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 5206)
  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: pCFJ1985 - [1-2] ENTR - Fluor - TagBFP2(PATC,
            ...
FEATURES             Location/Qualifiers
     source          1..5206
                     /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(2794..2893)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2910..2929)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2911..2929)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2934..2950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2934..2950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3052..3071)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3063..3872
                     /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(3130..3149)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4019..4608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4509..4528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4762..4779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4938..4965)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5057..5143)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5192..5206,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5206,1..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 gttttcttat gggaaagaag gaaaaaaccg agattttact
      601 tgaaaaattg aatttttcgc gggattttca ccaaaaattg ttgaatattc attatttcac
      661 gctgtaaaac aaaaaaaaaa aaaatcaaaa actacgttga aatcgcgttt ttaagcgaat
      721 tttcttcaga attgccagat tttaacccca aattttgcag tttttaaata aaatttcacc
      781 ttttcggctc aaattgtaga ttttcctgaa aatttagtac aaaaaacaat ttcctcgtaa
      841 atttttcaaa tagattttca gagggattca cctgggagcg tgtcaccacc tacgaggacg
      901 gaggagtcct caccgccacc caagacacct ccctccaaga cggatgcctc atctacaacg
      961 tcaagatccg tggagtcaac ttcacctcca acggaccagt catgcaaaag gtaagttatg
     1021 tctacctgcc tgcctaccgc ctaaattttt tgtgaagttt cttcaaaaaa tccagaaaaa
     1081 aaaacaattt tcatacgatt ttttccctta aaattgtgaa ttttcatgct ttttagcccc
     1141 aaaagtcatt atttgagaaa aaatttcata caaaaaagtt ttgagaaata cacaattttt
     1201 taaatgtaat tttcaaattt tcaattttca actagaaaat tcacaaaact tgtaaatttt
     1261 ggaccaaaac atttatacaa ttactttttt tgaatctaat aactacaata actacaaatt
     1321 taacattttc agaagaccct cggatgggag gccttcaccg agacgctcta cccagccgac
     1381 ggaggactgg agggacgtaa cgacatggcc ctcaagctcg tcggaggatc gcacctcatc
     1441 gccaacgcca agaccaccta ccgttccaag aagccagcca agaacctcaa gatgccagtc
     1501 tactacgtcg actaccgtct ggagcgtaag gtaagttctt tttttgaaaa gtgcagtttg
     1561 tagtctaatt ttcatttatt ttctttttaa aaaacgcatc aatttttaaa tattttggga
     1621 caaaatccga aaactgtact aatttgtagt ttgtaaaatt aaaaaaaaaa cgcaaaaaaa
     1681 tgttttcaaa aatgcttaga ataaaaataa aatttctaaa aaattgaaca aaataaacat
     1741 ttaaaaattc aaaaagtttg aaaaaatgca aagtttttta atgcaaaaaa tatttaatgt
     1801 ttaaacaaat ttaaaaaatg ttgtaagaat atgtttagag acagttttta aatgttaaat
     1861 tgcaaaaaaa aaaagcaaaa aaaaatttga aaaatgtaac agaaaatttt aattaaaaaa
     1921 gtcgtgtttt tttcaaacct gcaattcttt gtaattttat ttataaagag aaacaaatca
     1981 aattgtatat tacgaaaaac gatactaaaa ttcgaaaatt tgcggttttt tgcgtaaaaa
     2041 atacggttcc gtaattttca ggaggccaac aacgagacgt acgtcgagca acacgaggtc
     2101 gccgtcgccc gttactgcga cctcccatcc aagctcggac acaagctcaa ctcgggaact
     2161 tctacgactt cggtaacgtc atccggtact tccccagccg gctcccgtcg tcgtaaggta
     2221 agttcataga ttttgaaaaa aagtttaaga actgaaaaat ggaataaaaa tatttaagag
     2281 catttttaaa tgtaaaatta acaaaaaaag cgcctaagaa tgtttccaaa acagtaaaaa
     2341 aaaggtttaa aaaaatgcaa aaaaaaattt aagatgcttt taattacaca aaaaatgaaa
     2401 gtaaaaaaaa ttaaaatatt gaaaaatgta tatttgtttg aaaaggtaca tttttcaatg
     2461 caaaaaatgt ttctaaaaat taaaaaaata tgtttaaatt gtgtaaagtg taaattaaaa
     2521 aataaaatta tgaaaaagta ttgtttcata ctttttctaa attttggttg aaaaatttaa
     2581 aattccagct ttttattcac attatgtgtt ctaaaaatct ttccccaatt ttcgggaccg
     2641 ttttccagta tatttccatt tttcaaaaag atttttaact gaaattcatg ttttcaatgc
     2701 taaaaatcaa taaaaaaaga atttttcagg ccaacccaac caagctctcc gagaacgcca
     2761 agaagctcgc caaggaggtc gagaacggat ccaacccagc tttcttgtac aaagttggca
     2821 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2881 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2941 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     3001 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     3061 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     3121 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3181 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3241 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3301 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3361 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3421 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3481 acagcgatcg cgtatttcgc ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3541 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3601 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3661 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3721 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3781 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3841 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3901 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3961 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4021 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaaacc accgctacca
     4081 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4141 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4201 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4261 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4321 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4381 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4441 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4501 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4561 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4621 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4681 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4741 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4801 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4861 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     4921 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     4981 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     5041 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     5101 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     5161 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgtt
//
