LOCUS       sequence-141312-        5169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  AUTHORS   Frokjaer-Jensen C, Davis MW, Ailion M, Jorgensen EM
  TITLE     Improved Mos1-mediated transgenesis in C. elegans.
  JOURNAL   Nat Methods. 2012 Jan 30;9(2):117-8. doi: 10.1038/nmeth.1865.
  PUBMED    22290181
REFERENCE   2  (bases 1 to 5169)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-01-30- 30"; volume: "9"; 
            issue: "2"; pages: "117-8"
COMMENT     Sequence Label: pCFJ326 - operon for germline expression
FEATURES             Location/Qualifiers
     source          1..5169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..810
                     /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(68..87)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      957..1545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1446..1465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1699..1716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1875..1902)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1994..2080)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2129..2151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2143..2160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2144..2160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2231..2354)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2360..2683
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2846..2874)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3622..3642
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(4885..4980)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5017..5036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5018..5036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5041..5057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5041..5057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(join(5159..5169,1..9))
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
       61 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
      121 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      181 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
      241 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
      301 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      361 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
      421 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
      481 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
      541 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
      601 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
      661 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
      721 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
      781 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
      841 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
      901 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1081 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1321 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     1381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1621 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1681 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1741 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1801 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     1861 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     1921 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     1981 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2041 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     2101 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     2161 cttaagctcg ggccctgcag ctctagagct cgaattctac aggtcactaa taccatctaa
     2221 gtagttggtt catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt
     2281 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt
     2341 cttgtacaaa gtgggataaa tgcaaaatcc tttcaagcat tcccttcttc tctatcactc
     2401 ttctttcttt ttgtcaaaaa attctctcgc taatttattt gcttttttaa tgttattatt
     2461 ttatgacttt ttatagtcac tgaaaagttt gcatctgagt gaagtgaatg ctatcaaaat
     2521 gtgattctgt ctgatgtact ttcacaatct ctcttcaatt ccattttgaa gtgctttaaa
     2581 cccgaaaggt tgagaaaaat gcgagcgctc aaatatttgt attgtgttcg ttgagtgacc
     2641 caacaaaaag aggaaacttt attgtgccgc caagaaaaaa gtctcaggtt gtatatttat
     2701 tcatcttatt gaatctgcca tttcctcgtt tttgcgagtt tatatacctt ccaattttct
     2761 ttctattgta ttttcaactt ctaattttaa ttcagggaaa ctgcttcaac gcatcatgag
     2821 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     2881 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
     2941 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatgggtaa gtttaaacat
     3001 atatatacta actaaccctg attatttaaa ttttcagcca acacttgtca ctactttctg
     3061 ttatggtgtt caatgcttct cgagataccc agatcatatg aaacggcatg actttttcaa
     3121 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
     3181 ctacaagaca cgtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc
     3241 aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     3301 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca
     3361 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat
     3421 gattttacta actaactaat ctgatttaaa ttttcagaac ttcaaaatta gacacaacat
     3481 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     3541 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     3601 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     3661 tggcatggat gaactataca aatgcccggg ggatcggtgg agctccaccg gtggcggccg
     3721 ctctagaact agtccaccaa agccatctgc caagggagcc aagaaggccg ccaagaccgt
     3781 cgttgccaag ccaaaggacg gaaagaagag acgtcatgcc cgcaaggaat cgtactccgt
     3841 ctacatctac cgtgttctca agcaagttca cccagacacc ggagtctcct ccaaggccat
     3901 gtctatcatg aactccttcg tcaacgatgt attcgaacgc atcgcttcgg aagcttcccg
     3961 tcttgctcat tacaacaaac gctcaacgat ctcatcccgc gaaattcaaa ccgctgtccg
     4021 tttgattctc ccaggagaac ttgccaagca cgccgtgtct gagggaacca aggccgtcac
     4081 caagtacact tccagcaagt aaaataaccc gcaataggat tcttttatca tcgaaatttc
     4141 tgtagattac ttacttcaaa taatcatgca cacaggttgt ccccgttaat tctttccttc
     4201 aatctctttc tctctttcac ctagtctgtg ccatttcaat atgattgcac cccggaaaat
     4261 atcaaaaact tcacttaaca agtgatccca tattacaaaa atattcaaat tccgaaaatc
     4321 gaaagatgca tttccgtttt attttaccga attttgatgt ttttgtaatg tgtttaaacc
     4381 ccttttacaa ttaacttatt aatgttttgt tttactcatc tcacttcccc ttcttacttt
     4441 gtgcttattt tatgttttag gtttttttat tgtatcataa aactggaaat tttgaaatag
     4501 gcttctttct ccctaaactt ttttaacagg tactgtaact ggtacatatc ctctgcccca
     4561 ccccgctccc aactcgtctt tgtctgtttg tttatacgaa atacaatttt taacgagtat
     4621 gcgggtttct gttagcttca acaatacgat cattcaaatt ttaaaattaa gaaactataa
     4681 aagcacagta tttacgtgag ttgaaacaaa atgccttctc caaatttctt aacactgaaa
     4741 aattttcgga aaaaaagcag aaattactcc gcaaaaaatt aaggacatca ttccaatatt
     4801 tttttttact tatttgatga aaaattaata tattccaata aattaaaatg aattatcatc
     4861 tcaggaatac tgtagaatta ctttcaactt tattatacaa agttggcatt ataaaaaagc
     4921 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     4981 ggagctccat ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg
     5041 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     5101 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     5161 aggggtgtt
//
